Cardiomyocytes isolated selleck chem Axitinib from both GLP 1 receptor knockout mice and re spective wild type animals responded to lixisenatide with an increase in fractional shortening. Expression of genes involved in the GLP 1 incretin axis A local GLP 1 release may exist in the heart, potentially Inhibitors,Modulators,Libraries competing with lixisenatide on the GLP 1 receptor. There fore we analyzed tissues for the expression of the GLP 1 precursor peptide, glucagon, and converting and degrading proteases, PCSK1, PCSK2 and DPP4. Neither GCG, nor PCSK1 or PCSK2 expression could be observed in cardiac Inhibitors,Modulators,Libraries samples ruling out the existence of a local GLP 1 generation. In parallel we determined the expression of GLP 1 recep tor using two different specific Taqman primer assays span ning different exonintron within the known rat sequence.

Use of both primers assays indicated a similar strong expression in pancreas, isolated pancreatic islets and lung, but failed to amplify specifically any signal from car diac tissue and cardiomyocytes. It seems that all cardiac tis sues lack Inhibitors,Modulators,Libraries components of a local incretin system with Inhibitors,Modulators,Libraries DPP IV being the exception. Discussion Since the early 1990s GLP 1 peptides and analogs have been investigated for treating T2DM because of their ability to enhance glucose dependent insulin secretion. In addition, GLP 1 agonists are hypothesized to have effects on the cardiovascular system beyond glycaemic control, which may be exploited for therapeutic benefit. Cardiovascular disease is the leading cause of death in pa tients with type 2 diabetes. In a recent prospective cohort study of one million U. S.

adults, diabetes was associated with a twofold increase in the risk of death from ischemic heart disease. Concerns about the cardiovascular safety of anti diabetic drugs resulted in the FDA recommendation that the CV risk should be more thoroughly addressed dur ing drug development. As a consequence of this new guideline cardiovascular Inhibitors,Modulators,Libraries outcome studies are currently on going in more than 26,000 type 2 diabetic patients directly treated with GLP 1 receptor agonists and more than 40,000 treated with a DPP IV inhibitors which also may act indir ectly via GLP 1. The overall goal of our study was to dem onstrate in a pre clinical set of models cardioprotective effects of the GLP 1 analog lixisenatide, building a further fundament for clinical testing in patients with reduced car diac function. In an acute model of global cardiac ischemia reperfusion injury, lixisenatide treatment during the last 10 minutes of ischemia and during the whole reperfusion period signifi cantly reduced infarct size. The observed cardioprotective effect was not associated http://www.selleckchem.com/products/Bicalutamide(Casodex).html with a significant change in car diac hemodynamics, as assessed by rate pressure product RPP, and particularly coronary flow.

However, according to our result, the exact GO modules of alpha estradiol was mainly associated with lipid process, while monorden showed strong connec tion with sterol selleck Abiraterone process. The two different processes indi cated that Hsp90 should have a second pathway to con nect with AD. Compared with GO categories of alpha estradiol and monoden, fulvestrant had two additional categories, phosphoglyceride metabolic process and phospholipid biosynthetic process, listed. Based on these, we hypothesize that estrogen might act the Inhibitors,Modulators,Libraries phospholipid pathway designed to alleviate AD. Literature mining helped us reveal that estrogen reduced the risk of AD by anti Ab. Estrogen accelerated bAPP trafficking and pre cludesmaximal Ab generation within the TGN by mod ulating TGN phospholipid levels, particularly those of phosphatidylinositol.

Therefore, fulvestrant, as estrogen blocker might activate phospholipid Inhibitors,Modulators,Libraries pathway to acceler ate b amyloid and aggravate AD. Based on our analy Despite that both estrogen receptor and Hsp90 inhibitors are promis ing drug targets for AD, estrogen receptor is a much better Inhibitors,Modulators,Libraries candidate. The success of three cases between mouse models and human demonstrated that our cross species analysis method was able to assess animal models similarity to humans disease state. Inhibitors,Modulators,Libraries The main basis may be that orthologous genes were not only conserved at the sequence level and perform similar functions in different organisms, but also the corresponding gene expression patterns were conserved on a global level, especially between the human and mouse.

Our result that microarrays of cell response to molecules or drugs showed similarity across cell lines or tissues, to some extent, also explained why our approach was feasible to test mouse models. Nevertheless, as the diabetes drug case showed, it should be noticed that intrinsic Inhibitors,Modulators,Libraries differ ences always existed in normal and pathobiology states between species. Therefore, it was sometimes not appropriate for an animal model to mimic human dis eases or drug response. Our method based on the analy sis of the relationship of function known drugs and human diseases utilizing microarray expression data per formed well in both situations. Additionally, due to the introduction of GO annotations and the application of the statistical analysis, this cross species method was able to provide bidirectional relationship between drugs and disease, and more clues about potential biological mechanisms.

By contrast, the distance method seemed to be oversimplified and couldnt provide such information. In this paper, we presented data only from mouse ani mal models, for the mouse was the main model for common human diseases. At present, the National Cen ter for Biotechnology Information Gene Expres sion Omnibus enrolled 1295 datasets on homo selleck kinase inhibitor sapiens and 1069 datasets on Mus musculus.

Immunoprecipitation Two five hundred ug of cell lysate proteins were incu bated with 4 ug of antibody overnight selleck chemicals Dorsomorphin on a rotator at 4 C. Recombinant Protein A/G ultra link resin or TrueblotW anti light chain IP beads were washed and added at 1 10 ratio of beads to lysate volumes, then mixed further for 2 3 hours at 4 Inhibitors,Modulators,Libraries C. Immunoprecipitation mixtures were Inhibitors,Modulators,Libraries microcentrifuged for thirty seconds, the beads washed, then pellets resuspended in 20 65 ul 2x sample loading buffer, boiled, cooled, and microcentrifuged before loading 10 15 ul into SDS PAGE gels. si RNA transfection Lyn siRNA and negative control siRNA were diluted to 250nM in antibiotic free OPTI MEM with Glutamax and mixed with an equal volume of trans fection reagent then incubated 20 minutes at room temperature with shaking before 1.

0 ml of each mixture was added to cells adhered to duplicate wells of a 6 well plate. An other 1. 0 ml of OPTI MEM containing 10% FBS but no antibiotics was added after 4 6 hours at 37 C, then the plates were incubated for 48, 72, 96, and 144 hours as noted. The kinetics and effectiveness of Lyn siRNA knock down was confirmed Inhibitors,Modulators,Libraries by Western blotting with anti Lyn or anti phospho Lyn. The protocol to determine the effect of Lyn siRNA knock down on Calu3 cell via bility was modified to ten replicate wells in 96 well plates of Calcein AM assay as described above. Results Constitutive phosphorylation of EGFR in NSCLC cell lines Constitutive phosphorylation of EGFR at Y 845 in Calu3 and H1975 cell lines, and at Y 992 was seen in Calu3, H1975, and A549 cell lines.

CLL cells did not express EGFR and nonspecific staining with anti phospho EGFR antibodies was not observed. PCR and SSCP assays did not detect activating mutations in Calu3 cells in exons 19 and 21 of the erbB1 gene, hence Calu3 served as the target of our investigations. H1975 cells on the other hand contain an activating muta tion in exon 21 resulting in Inhibitors,Modulators,Libraries EGFR phosphorylation. To investigate mechanisms of constitutive activation of EGFR, autophosphorylation was inhibited with EGFR tyrosine kinase inhibitor AG1478, and later confirmed with erlotinib. Phosphorylation of Y 992 and Y 845 of EGFR were still detectable in unstimulated, serum starved Calu3 cells confirming that they are not auto phosphorylation sites, but are phosphorylated by up stream kinases. AG1478 was functional as it inhibited down stream phosphorylation of Akt.

Ligands were not responsible for constitutive phosphorylation of EGFR in unstimulated, serum starved Calu3 cells as increments of EGF neutralizing monoclonal antibody, LA1, from 12. 5 to 50 ug/ml failed to inhibit phosphoryl ation. LA1, binds the EGFR extracellular domain and competes for binding with Inhibitors,Modulators,Libraries ligands. EGF, TGF, and AR. www.selleckchem.com/products/Gemcitabine-Hydrochloride(Gemzar).html LA1 was effective as it inhibited EGF ligand induced Y 992 and Y 845 phosphorylation in H1975 cells.

It has been reported that both gefitinib and its des methyl metabolite formed through CYP2D6, inhib this website ited with a similar potency and selectivity subcellular EGFR tyrosine kinase activity. However, M3 was 15 times less active in a cell based assay and consequently it was assumed that this metabolite was unlikely to con tribute to the activity of gefitinib in vivo due to poor cell penetration. On the contrary, when metabolites M1, M2 and M3 were tested in our responsive cell models at concentra tions equivalent to that of gefitinib, they exhibited a signif icant inhibition of EGFR autophosphorylation and proliferation in intact cells, indicating their ability to enter cells and to interact with the catalytic domain of EGFR.

Finally, in gefitinib resistant cell lines M1, M2 and M3 metabolites were poorly effective indicating that at least these metabolites did not produce additive toxic effects in NSCLC cell lines. In contrast to its abundant hepatic expression, CYP3A4 seems to play a minor role in lung metabolism, being expressed in only about 20% Inhibitors,Modulators,Libraries of cases. Real time PCR analysis confirmed the lack of expression of this isoform in our NSCLC cell models, as reported for A549 cells. CYP2D6 was detected in all cell lines, whereas both CYP1A1 and CYP1A2 were expressed at significant levels in sensitive cells. Inducibility of CYP1A1 and CYP1A2 transcripts by gefitinib was clearly demonstrated in sensitive cell lines, while induction of CYP1A1 mRNA was not detected in resistant cell lines. EROD activity demonstrated a 3 6 fold induction of CYP1A1 elicited by gefitinib in sensitive cells.

To the best of our knowledge, this is the first time that the Inhibitors,Modulators,Libraries induction by gefitinib Inhibitors,Modulators,Libraries of relevant metabolic enzyme has been demonstrated. Inhibitors,Modulators,Libraries The reason why gefitinib induces CYP expression Inhibitors,Modulators,Libraries and activity only in sensitive cells could be ascribed to the ability of gefitinib to inhibit signal transduction pathway downstream EGFR. It has been recently demonstrated that EGF represses the dioxin mediated induction of CYP1A1 in normal human keratinocytes preventing recruitment of the p300 coactivator. Therefore, EGFR signalling is a repressor of the aryl hydrocarbon receptor and regulates the transcription of numerous genes including CYP1A1. In this context, EGFR inhibi tors such as gefitinib, erlotinib, lapatinib or cetuximab might affect the induction of CYP1A1 in those cell types in which the drug effectively inhibits signalling controlled by EGFR.

The inhibition of MAPK pathway might repre sent a link between EGFR inhibition and CYP1A1 induc tion since PD98059 and U0126, well known MEK1/2 inhibitors, induced CYP1A1 activity as did gefitinib in H322 cells, while none of PI3K/AKT/mTOR selleck chem inhibitor inhibitors tested was effective. It is noteworthy that constitutive activation of signaling pathways downstream of EGFR is a recognized mechanism or resistance against reversible EGFR tyrosine kinase inhibitors.

Therefore, we validated the methylation and resul tant decreased expression of BMX and SOX1 in the non invasive cells. Functional role of Bmx and Sox1 during invasion To further determine the role of Bmx and Sox1 during the process of invasion we performed the invasion assay with DU145 cells stably infected with shRNAs directed against Sox1or different Bmx. A significant decrease in expression of SOX1 and BMX following induction with 1 ug/mL of doxycycline for 24 hours was first verified using western blotting. Upon induction with Dox, the shRNA is turned on and a downstream red fluorescent protein demonstrates efficiency of this induction. Densitometry analysis was per formed to compare expression of individual clones with the NS cells, and no significant differences in protein expression were seen using the non silencing con trols.

In addition, SOX1 shRNA cells demonstrated a significant decrease in proliferation compared to either the parental Inhibitors,Modulators,Libraries cell line or the NS infected line, as well as a significant decrease in invasion toward SCM. However, there was not a significant difference using the shBMX lines, except for a slight reduction in invasion using clone 3. Interestingly, a small increase in proliferation Inhibitors,Modulators,Libraries was seen with the shBMX clones. Further promoter tiling array analysis using two short term cultures primary prostate tumor cell lines, PCSC1 and PCSC2, determined that Sox1, and not Bmx, was methylated in the invasive population of cells. Overall, we demonstrate that Sox1is differentially methylated Inhibitors,Modulators,Libraries within the invasive CSC population and the shRNA studies indicate it could be selectively targeted to block invasion.

Role of SOX1 during differentiation In addition to the method presented here, prostate TICs can also be isolated by culturing total cells Inhibitors,Modulators,Libraries in SCM where structures called prostato spheres are generated. The prostatospheres are multicellular globes that develop from cells that sur vive anchorage independent conditions in vitro, and are frequently used when analyzing the ability of TICs to self renew or differentiate upon the addition of serum. Using this assay as a model, a greater number of prosta tospheres were isolated from DU145 NS cells compared to shSOX1 cells. When invasive DU145 cells were isolated and cultured in SCM, prostatospheres were maintained for up to 3 passages and if these cells were further cultured in the presence of 1% human serum, the vector control cells rapidly differentiated and proliferated, while the shSOX1 cells did not.

These observations suggest that not only does Sox1 play a role in regulating invasion, but it can also regulate the maintenance of stem ness in culture. Ingenuity pathway analysis defines pathways of differentially Inhibitors,Modulators,Libraries methylated genes within invasive sub populations of cells Each data set of differentially methylated genes was then extracted leave a message and uploaded to the Ingenuity server to identify common gene pathways that are regulated during the process of invasion.

On the contrary, overexpression of Aur A led to Akt activation and I?Bdown regulation, subsequently induced NFB p65 nuclear translocation to enhance cell survival. Moreover, suppression of Akt by either API 2 or siAkt prevented Aur A induced I?Breduc tion and Bcl xL elevation. Thus, our data demonstrated http://www.selleckchem.com/products/U0126.html that Aur A downregulated I?Bvia Akt activation, trigger ing NFB p65 nuclear translocation, and subsequently activating target gene Bcl xL to promote survival in cancer cells. Inactivation of PTEN leads to constitutively activate PI3K/ Akt pathway. Recently, Aur A was found to abrogate the DNA binding and transactivation activity of p53 and sub sequently inhibit its downstream target genes including PTEN by phosphorylating Ser 215. PTEN expression was significantly reduced in Aur A overexpressed cells with activated Akt activity.

Here, Inhibitors,Modulators,Libraries we showed that overexpression Inhibitors,Modulators,Libraries of Aur A increased the phosphorylation of Akt at Ser 473. Consistently, previous report showed that Aur A activated Akt in a p53 dependent man ner to induce cell Inhibitors,Modulators,Libraries survival and chemoresistance in ovarian cancer cells. Thus, it is conceivable that Aur A activates Akt via inhibiting PTEN. Akt promotes cell survival by its ability to phosphorylate and inactivate several pro apoptotic targets including GSK 3. We showed that inhibition of Aur A resulted in suppressed phosphorylation of both Akt and GSK 3, according with one recent study that Aur A promoted cell proliferation by increasing the phosphorylation of GSK 3?.

On the other hand, another work reported that Akt inhibitor A 443654 interfered with mitotic progres sion by decreasing Aur A expression, suggesting Akt acts upstream of Aur A by regulating its transcription level. We and others showed that Aur A contributed to cell survival, chemoresistance and migration Inhibitors,Modulators,Libraries via activation of Akt, suggesting a positive feedback interplaying between Aur A and Akt. Akt plays a part in activation of NFB signaling pathway and exerts a positive effect on NFB function by phos phorylation and activation of IKK, a kinase that phospho rylates and induces proteolytic degradation of the NFB inhibitor, I?B.Interestingly, several recent reports have suggested that Aur A kinase may serve both upstream and downstream of the IKK complex components. IKK complex includes two catalytic components, IKKand IKK?.

As a downstream target, Aur A was phos phorylated by IKKat threonine residue 288, a site which is important for its kinase activity. Depletion Inhibitors,Modulators,Libraries of IKK resulted in the up regulation of Aur A protein, and IKK functioned as an antagonist of Aur A signaling during mitosis in normal cells. On the other hand, we showed that Aur A promoted cell survival through acti vated IKK/NFB signaling pathway, consistent with pre vious kinase inhibitor Enzalutamide reports. Thus, there may be a reciprocal regulation between Aur A and IKK complex.

Continuous Carfilzomib buy accumulation of Imatinib Mesylate structure both PDGF and VEGF was observed in all Inhibitors,Modulators,Libraries the HMEC lines. Even so, our results suggest that the regulatory mechanisms that control the secretion of these two proteins are distinct. In HMEC, secretion of PDGF seems to be a constitutive process that is independent of Erk and PI3K/Akt pathways. during In contrast, synthesis and secretion of VEGF Inhibitors,Modulators,Libraries is a direct and spontaneous response to HER signaling through EGF Inhibitors,Modulators,Libraries stimulation, and both MAPK/Erk and PI3K/Akt path ways appear Inhibitors,Modulators,Libraries to have a role in VEGF secretion. Our results are consistent with previous studies that demon strate that HER1 regulates VEGF expression in a variety Inhibitors,Modulators,Libraries of different cancer cell lines.

Conclusion In summary, we examined the interaction Inhibitors,Modulators,Libraries between HER receptor levels and protein secretion in HMEC, with the Inhibitors,Modulators,Libraries dual goals of gaining insight into the molecular pro cesses that regulate Inhibitors,Modulators,Libraries the secretion of proteins that influ ence cancer development and progression, and therefore may serve as biomarkers that can be used to assess molecular events in tumor tissue. By studying the secre tion patterns of a panel of proteins that are associated with a variety of human cancers, we gained novel insight into how HER activation results Inhibitors,Modulators,Libraries in a concerted change in multiple classes of secreted proteins that not only alter the cellular microenvironment through matrix reconstruction, angiogenesis, and paracrine interactions with stromal and immune cells, but also provide auto crine feedback to the tumor epithelial cells that results in further enhanced growth and migration.

Therefore, this study Inhibitors,Modulators,Libraries suggests that changes in protein secretion may be an important factor by which HER activation stimulates mammary tumor formation. Effects of sEHi on FAS and IL 6 protein levels in PBMCs t AUCB had a dose dependent Inhibitors,Modulators,Libraries inhibitory effect on Inhibitors,Modulators,Libraries the expression of FAS,IL 6 protein in PBMCs from the ACS group. Compared with the control group,levels of 10,50,100 umol L t AUCB had inhib ited the expression of FAS,IL 6 protein in PBMCs from the ACS Inhibitors,Modulators,Libraries group. The relative expression levels of FAS protein in the t AUCB 0,10,50 and Inhibitors,Modulators,Libraries 100 umol L groups were respectively.

The relative expression of IL 6 protein in the t AUCB 0,10,50 and 100 umol L groups were respectively.

selleck chem inhibitor Discussion Numerous studies have demonstrated that the inflamma Inhibitors,Modulators,Libraries tory level is increased in the ACS patients. Shantsila and Lip highlighted that Enzastaurin order monocytes were actively involved in the pathological processes related to ACS,which promoted the synthesis of pro inflammatory molecules,such as IL 6,tumor www.selleckchem.com/products/Y-27632.html necrosis factor and hs CRP. Among them,hs CRP has been proved to be the strongest and most significant predictor of the in flammatory level and the risk of plaque instability and rupture.

2 ��M Wortmannin, 25 ��M PD98059, 10 ��M U0126. Stock Enzastaurin purchase solutions of each reagent SKI 606 selleck compound were Inhibitors,Modulators,Libraries individually prepared in DMSO and the final concentration of DMSO in the culture medium was 0. 1% in all treatments, includ ing controls. At least two independent experiments were performed, with five biological replicates in each experiment, for all results described here. Samples were stored at 80 C until analysis. ELISA Microarray Analysis Concentrations of individual proteins in CM and cell lysates were quantitatively measured using sandwich enzyme linked immunosorbent assay, as pre viously described in detail. Briefly, ELISA microarray chips were custom manufactured using aminosilanated, 25×75 mm glass slides stamped with a hydrophobic barrier that was used to cre ate 16 wells on each slide.

Inhibitors,Modulators,Libraries The ELISA reagents used in these analyses have been previously evaluated and shown to have no cross reactivity Inhibitors,Modulators,Libraries and to be able to quantita Inhibitors,Modulators,Libraries tively detect purified antigens that were spiked into human Inhibitors,Modulators,Libraries serum. The capture antibodies were sus pended in PBS at concentrations ranging from 0. 5 to 1. 0 mg/ml. These antibodies were printed using a GeSiM noncontact NanoPlotter NP2 printer. Sixteen identical chips were printed on each slide, such that each chip was isolated by a hydrophobic barrier. Each capture antibody and control reagent was printed in quadruplicate on each chip. Successful printing was confirmed using the RedReflect capability on the ScanArray ExpressHT laser scanner.

Inhibitors,Modulators,Libraries The printed slides were blocked with 1% casein in PBS at 4 C and, Inhibitors,Modulators,Libraries after washing, stored desiccated and under vacuum at 20 C until use.

Inhibitors,Modulators,Libraries In order to generate standard curves Inhibitors,Modulators,Libraries for each of the ELISA analyses, a single mixture containing all the anti gens was prepared in 0. 1% casein in PBS and containing 100 pg/ml green fluorescent protein. This stock solution of the standard mixture was aliquoted and stored at 80 C, and an aliquot was thawed on a daily Inhibitors,Modulators,Libraries basis for Inhibitors,Modulators,Libraries each ELISA microarray analyses. For this analysis, the stan dard stock solution was serially diluted 3 fold to create at least 7 dilutions of the standard mixture. Each dilu tion, and an antigen free blank, was analyzed on three seperate chips.

Several Inhibitors,Modulators,Libraries incubation steps are included in processing the ELISA microarray chips.

Washes were performed between each incubation step by submerging Inhibitors,Modulators,Libraries the Inhibitors,Modulators,Libraries slides in PBS containing 0. 05% Tween 20.

Either 20 ul of the standard mixture or an individual, diluted sam ple was then pipetted onto each of three replicate chips. Samples were then incubated overnight in a closed chamber with saturated humidity and gentle mixing. Slides were then incubated Inhibitors,Modulators,Libraries with a cocktail of all Calcitriol mechanism detec tion antibodies sellckchem selleck Imatinib Mesylate in 0. 1% casein/PBS buffer for 2 h. The level of biotinylation was then increased using the bioti nyltyramide amplification system. Finally, slides were submerged in 1 ug/ml of Cy3 or Alexa 647 conjugated streptavidin in PBS T, and incubated for 1 h in dark.

Preparation of compounds for screening The Maybridge Hitskit 3000 library con sists of 3000 chemically diverse compounds. The library was delivered find more info in 36 racks each containing 80 compounds dissolved in DMSO to 10 mg/ml. For the screening, ali quots of the DMSO solutions were transferred to 96 well plates and were further diluted with PBS to Inhibitors,Modulators,Libraries obtain stock solutions of 100 ug/ml from which four different 384 well plates for screening were prepared with final test concen trations of 1 ug/ml. In all steps, the Biomek 2000 pipetting station connected to a plate stacker carousel in a safety cabinet was used. For dose response studies, plates containing VLX40 and other compounds were prepared by 10 fold serial dilu tions in the concentrations 0. 004 to 40 uM using the same robotic system.

The plates were stored at 70 C until further use. The screening identified one compound with higher activity against 8226/Dox40 cells compared to its parental counterpart RPMI 8226. This compound, chem Inhibitors,Modulators,Libraries ically a quinoline alkaloid, was designated VLX40, and subjected for detailed studies. Measurement of cancer Inhibitors,Modulators,Libraries drug activity The Fluorometric Microculture Cytotoxicity Assay, FMCA, described in detail previously, was used for measure ment of the cytotoxic effect of library compounds and the established standard drugs. The FMCA is based on measurement of fluorescence generated from hydrolysis of fluorescein diacetate to fluorescein by cells with intact plasma membranes. Cells were seeded in the drug prepared 384 well plates using the pipetting robot Pre cision 2000.

The number of cells per well was 2,500 5,000 for solid tumor samples and 10,000 20,000 for leukemic samples. In each plate, two columns without drugs served as controls and Inhibitors,Modulators,Libraries one column with medium only served as blank. The plates were incubated for 72 h and then transferred to an integrated HTS SAGIAN Inhibitors,Modulators,Libraries Core System consisting of an ORCA robot with CO2 incubator, dispenser module, washer module, de lidding station, plate hotels, barcode reader, liquid handler and a multipurpose reader for automated FMCA. Quality criteria for a successful assay included a mean coefficient of variation of less than 30% in the control wells and a fluorescence signal in control wells of more than 5 times the blank. Survival index is defined as the fluorescence of test wells in percentage of controls with blank values subtracted.

Multiparametric or high content evaluation of apoptosis and cell cycle arrest The fluorescence microscope ArrayScan High Content Screening system was used to study apoptosis and cell cycle arrest. For these assays, cells were seeded into 96 well plates, left to attach over night, before test compounds were added. Cell death characteristics were studied using a multi parametric HCS assay described in detail previously. Apoptosis was evaluated after 6, 24 and 48 h exposure to VLX40 in MCF 7 cells.