We discovered a highly potent ATP-competitive PI3K/Vps34 dual inhibitor, PI3KD/V-IN-01, which displayed 10-1500 fold selectivity over additional PI3K isoforms and did not inhibit some other kinases in the kinome. PI3KD/V-IN-01 than the FLT3 wt expressing cells. In AML cell inoculated xenograft mouse model, PI3KD/V-IN-01 exhibited dose-dependent anti-tumor growth efficacies. These results suggest that dual inhibition of PI3K and Vps34 might be a useful approach to improve the PI3K inhibitor’s anti-tumor effectiveness. and IP kinase assay shown only moderate inhibition of mTORC1 kinase by PI3KD/V-IN-01 (EC50 of 1700 nM), which is much less potent than activity displayed against PI3K and vps34 kinases. (Supplemental Number 3) In HeLa cells, PI3KD/V-IN-01 efficiently prevented LC3BII build up in the presence of EBSS (Earle’s balanced salt remedy) and HCQ (hydrochloroquine) with an EC50 of 413 nM. (Number ?(Figure1E)1E) In addition, an immunofluorescence experiment showed that PI3KD/V-IN-01 increased LC3B puncta AG-120 in HeLa cells inside a dose-dependent way, which was similar to the Vps34 specific inhibitor Vps34-IN-1, but not for the PI3K inhibitor CAL-101 or pan-PI3K inhibitor GDC-0941. (Number ?(Number1F1F and Supplemental Number 2) This is not amazing as HeLa cells express Vps34 but not PI3K. In accordance with the founded function of Vps34 in membrane trafficking [16, 17], the lysosome marker Light1 localization was affected by PI3KD/V-IN-01 and Vps34-IN-1, however not GDC-0941 or CAL-101. (Number ?(Figure1F)1F) These biochemical and cellular data demonstrate that PI3KD/V-IN-01 was a highly potent and selective PI3K/Vps34 dual inhibitor. Open in a separate window Number 1 Biochemical and pharmacological characterization of PI3KD/V-IN-01A. Chemical structure of PI3KD/V-IN-01. B. ADP-Glo? Biochemical IC50 dedication of PI3KD/V-IN-01 against a panel of PI3K-related kinases. C. Dedication of the inhibitory effect of PI3KD/V-IN-01 against class I PI3Ks in the cellular context. Specifically, PI3K in NIH-3T3 cells with PDGF-BB activation; PI3K in NIH-3T3 cells with LPA activation; PI3K in Natural264.7 cells with C5a activation; PI3K in Raji cells with anti-IgM activation. D. Selectivity profile of PI3KD/V-IN-01 in the DiscoveRx’s KinomeScan? platform. E. Effect of PI3KD/V-IN-01 on autophagy in HeLa cells using co-culture of EBSS and HCQ (25 M) and investigating LC3BII manifestation. F. Immuno-fluorescent imaging analysis of the effect of PI3K inhibitors on LC3BII manifestation in HeLa cells and of Light1 manifestation in HeLa cells treated with AG-120 PI3K inhibitors. Table 1 Quantification of PI3KD/V-IN-01 EC50 against class I PI3Ks IP kinase assay. Activity of the downstream AKT kinase mediator, GSK3, and downstream transcription regulator, NFB, were not affected in any of the cell lines. What’s noteworthy is certainly that in MEC-2 cells, benefit was inhibited by PI3KD/V-IN-01, CAL-101 and GDC-0941, but this sensation was not seen in OCI-AML-2, MV4-11 and OCI-AML-3 cells. These outcomes demonstrate that PI3K is certainly inhibited in the mobile framework successfully, nevertheless differential responsiveness of specific signaling molecules towards the inhibitors examined claim that these cells might depend on different hereditary/signaling network backgrounds. Open up in another window Body 2 Aftereffect of PI3KD/V-IN-01 on mobile signaling, cell routine development, and autophagyA. Aftereffect of PI3KD/V-IN-01 on PI3K- mediated signaling pathways in OCI-AML-2 (AML), MV4-11 (AML), OCI-AML-3(AML) and MEC-2 (CLL) cell lines. B. Apoptotic aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, MEC-2 and OCI-AML-3 cells. C. Autophagy interruption aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. D. Cell routine progression aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. We following examined the result of PI3KD/V-IN-01 on induction of apoptosis. Oddly enough, limited to MV4-11 was obvious PARP cleavage noticed, which implies drug-induced apoptosis of the line clearly. (Body ?(Figure2B)2B) For OCI-AML-2 and MEC-2, there is only a humble induction of apoptosis (noticeable at 3 M to 10 M), as well as for OCI-AML-3, zero apoptosis was noticed, at concentrations up to 10 M even. Of relevance, a concentration-dependent upsurge in degrees of the autophagy markers, P62 and LC3BII, was seen in OCI-AML-2, MEC-2 and OCI-AML-3. (Body ?(Figure2C)2C) However, this phenomenon had not been noticed for MV4-11 cell line. Cell routine analysis demonstrated that after 48 h treatment with PI3KD/V-IN-01, MV4-11, MEC-2 and OCI-AML-3 cells could possibly be imprisoned in the G0/G1 stage, while there is no obvious cell routine inhibition noticed for drug-treated OCI-AML-2 cells. (Body ?(Figure2D2D) Mix of CAL-101 and Vps34-IN-1 recapitulates the dual inhibitory aftereffect of PI3KD/V-IN-01 We following investigated set up mix of the selective PI3K inhibitor, CAL-101, and.Cell proliferation were determined after treatment using the substances for 72 hours. PI3K and Vps34 may be a good method of enhance the PI3K inhibitor’s anti-tumor efficiency. and IP kinase assay confirmed only humble inhibition of mTORC1 kinase by PI3KD/V-IN-01 (EC50 of 1700 nM), which is a lot less powerful than activity shown against PI3K and vps34 kinases. (Supplemental Body 3) In HeLa cells, PI3KD/V-IN-01 successfully prevented LC3BII deposition in the current presence of EBSS (Earle’s well balanced salt alternative) and HCQ (hydrochloroquine) with an EC50 of 413 nM. (Body ?(Figure1E)1E) Furthermore, an immunofluorescence experiment showed that PI3KD/V-IN-01 improved LC3B puncta in HeLa cells within a dose-dependent method, which was like the Vps34 particular inhibitor Vps34-IN-1, however, not for the PI3K inhibitor CAL-101 or pan-PI3K inhibitor GDC-0941. (Body ?(Body1F1F and Supplemental Body 2) This isn’t astonishing as HeLa cells express Vps34 however, not PI3K. Relative to the set up function of Vps34 in membrane trafficking [16, 17], the lysosome marker Light fixture1 localization was suffering from PI3KD/V-IN-01 and Vps34-IN-1, nevertheless not really GDC-0941 or CAL-101. (Body ?(Figure1F)1F) These biochemical and mobile data demonstrate that PI3KD/V-IN-01 was an extremely powerful and selective PI3K/Vps34 dual inhibitor. Open up in another window Body 1 Biochemical and pharmacological characterization of PI3KD/V-IN-01A. Chemical substance framework of PI3KD/V-IN-01. B. ADP-Glo? Biochemical IC50 perseverance of PI3KD/V-IN-01 against a -panel of PI3K-related kinases. C. Perseverance from the inhibitory aftereffect of PI3KD/V-IN-01 against course I PI3Ks in the mobile context. Particularly, PI3K in NIH-3T3 cells with PDGF-BB arousal; PI3K in NIH-3T3 cells with LPA arousal; PI3K in Organic264.7 cells with C5a arousal; PI3K in Raji cells with anti-IgM arousal. D. Selectivity account of PI3KD/V-IN-01 in the DiscoveRx’s KinomeScan? system. E. Aftereffect of PI3KD/V-IN-01 on autophagy in HeLa cells using co-culture of EBSS and HCQ (25 M) and looking into LC3BII appearance. F. Immuno-fluorescent imaging evaluation of the result of PI3K inhibitors on LC3BII appearance in HeLa cells and of Light fixture1 appearance in HeLa cells treated with PI3K inhibitors. Desk 1 Quantification of PI3KD/V-IN-01 EC50 against course I PI3Ks IP kinase assay. Activity of the downstream AKT kinase mediator, GSK3, and downstream transcription regulator, NFB, weren’t affected in virtually any from the cell lines. What’s noteworthy is certainly that in MEC-2 cells, benefit was considerably inhibited by PI3KD/V-IN-01, GDC-0941 and CAL-101, but this sensation was not seen in OCI-AML-2, OCI-AML-3 and MV4-11 cells. These outcomes demonstrate that PI3K is certainly successfully inhibited in the mobile context, nevertheless differential responsiveness of specific signaling molecules towards the inhibitors examined claim that these cells might depend on different hereditary/signaling network backgrounds. Open up in another window Body 2 Aftereffect of PI3KD/V-IN-01 on mobile signaling, cell routine development, and autophagyA. Aftereffect of PI3KD/V-IN-01 on PI3K- mediated signaling pathways in OCI-AML-2 (AML), MV4-11 (AML), OCI-AML-3(AML) and MEC-2 (CLL) cell lines. B. Apoptotic aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. C. Autophagy interruption aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. D. Cell routine progression aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. We following examined the result of PI3KD/V-IN-01 on induction of apoptosis. Oddly enough, limited to MV4-11 was obvious PARP cleavage noticed, which obviously suggests drug-induced apoptosis of the line. (Shape ?(Figure2B)2B) For OCI-AML-2 and MEC-2, there is only a moderate induction of apoptosis (apparent at 3 M to 10 M), as well as for OCI-AML-3, zero apoptosis was noticed, sometimes at concentrations up to 10 M. Of relevance, a concentration-dependent upsurge in degrees of the autophagy markers, LC3BII and p62, was seen in OCI-AML-2, OCI-AML-3 and MEC-2. (Shape ?(Figure2C)2C) However, this phenomenon had not been noticed for MV4-11 cell line. Cell routine analysis demonstrated that after 48 h treatment with PI3KD/V-IN-01, MV4-11, OCI-AML-3 and MEC-2 cells could possibly be caught in the G0/G1 stage, while there is no obvious cell routine inhibition noticed for drug-treated OCI-AML-2 cells. (Shape ?(Figure2D2D) Mix of CAL-101 and Vps34-IN-1 recapitulates the dual inhibitory aftereffect of PI3KD/V-IN-01 We following investigated set up combination.Hematoxylin and eosin staining of cell and cells areas. FLT3 wt expressing cells. In AML cell inoculated xenograft mouse model, PI3KD/V-IN-01 exhibited dose-dependent anti-tumor development efficacies. These outcomes claim that dual inhibition of PI3K and Vps34 may be a good method of enhance the PI3K inhibitor’s anti-tumor effectiveness. and IP kinase assay proven only moderate inhibition of mTORC1 kinase by PI3KD/V-IN-01 (EC50 of 1700 nM), which is a lot less powerful than activity shown against PI3K and vps34 kinases. (Supplemental Shape 3) In HeLa cells, PI3KD/V-IN-01 efficiently prevented LC3BII build up in the current presence of EBSS (Earle’s well balanced salt option) and HCQ (hydrochloroquine) with an EC50 of 413 nM. (Shape ?(Figure1E)1E) Furthermore, an immunofluorescence experiment showed that PI3KD/V-IN-01 improved LC3B puncta in HeLa cells inside a dose-dependent method, which was like the Vps34 particular inhibitor Vps34-IN-1, however, not for the PI3K inhibitor CAL-101 or pan-PI3K inhibitor GDC-0941. (Shape ?(Shape1F1F and Supplemental Shape 2) This isn’t unexpected as HeLa cells express Vps34 however, not PI3K. Relative to the founded function of Vps34 in membrane trafficking [16, 17], the lysosome marker Light1 localization was suffering from PI3KD/V-IN-01 and Vps34-IN-1, nevertheless not really GDC-0941 or CAL-101. (Shape ?(Figure1F)1F) These biochemical and mobile data demonstrate that PI3KD/V-IN-01 was an extremely powerful and selective PI3K/Vps34 dual inhibitor. Open up in another window Shape 1 Biochemical and pharmacological characterization of PI3KD/V-IN-01A. Chemical substance framework of PI3KD/V-IN-01. B. ADP-Glo? Biochemical IC50 dedication of PI3KD/V-IN-01 against a -panel of PI3K-related kinases. C. Dedication from the inhibitory aftereffect of PI3KD/V-IN-01 against course I PI3Ks in the mobile context. Particularly, PI3K in NIH-3T3 cells with PDGF-BB excitement; PI3K in NIH-3T3 cells with LPA excitement; PI3K in Natural264.7 cells with C5a excitement; PI3K in Raji cells with anti-IgM excitement. D. Selectivity account of PI3KD/V-IN-01 in the DiscoveRx’s KinomeScan? system. E. Aftereffect of PI3KD/V-IN-01 on autophagy in HeLa cells using co-culture of EBSS and HCQ (25 M) and looking into LC3BII manifestation. F. Immuno-fluorescent imaging evaluation of the result of PI3K inhibitors on LC3BII manifestation in HeLa cells and of Light1 manifestation in HeLa cells treated with PI3K inhibitors. Desk 1 Quantification of PI3KD/V-IN-01 EC50 against course I PI3Ks IP kinase assay. Activity of the downstream AKT kinase mediator, GSK3, and downstream transcription regulator, NFB, weren’t affected in virtually any from the cell lines. What’s noteworthy can be that in MEC-2 cells, benefit was considerably inhibited by PI3KD/V-IN-01, GDC-0941 and CAL-101, but this trend was not seen in OCI-AML-2, OCI-AML-3 and MV4-11 cells. These outcomes demonstrate that PI3K can be efficiently inhibited in the mobile context, nevertheless differential responsiveness of particular signaling molecules towards the inhibitors examined claim that these cells might depend on different hereditary/signaling network backgrounds. Open up in another window Shape 2 Aftereffect of PI3KD/V-IN-01 on mobile signaling, cell routine development, and autophagyA. Aftereffect of PI3KD/V-IN-01 on PI3K- mediated signaling pathways in OCI-AML-2 (AML), MV4-11 (AML), OCI-AML-3(AML) and MEC-2 (CLL) cell lines. B. Apoptotic aftereffect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. C. Autophagy interruption effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. D. Cell cycle progression effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. We next examined the effect of PI3KD/V-IN-01 on induction of apoptosis. Interestingly, only for MV4-11 was apparent PARP cleavage observed, which clearly suggests drug-induced apoptosis of this line. (Figure ?(Figure2B)2B) For OCI-AML-2 and MEC-2, there was only a modest induction of apoptosis (evident at 3 M to 10 M), and for OCI-AML-3, no apoptosis was observed, even at concentrations up to 10 M. Of relevance, a concentration-dependent increase in levels of the autophagy markers, LC3BII and p62, was observed in OCI-AML-2, OCI-AML-3 and MEC-2. (Figure ?(Figure2C)2C) However, this phenomenon was not observed for MV4-11 cell line. Cell cycle analysis showed that after 48 h treatment with PI3KD/V-IN-01, MV4-11, OCI-AML-3 and MEC-2 cells could be arrested in the G0/G1 phase, while there was no apparent cell cycle inhibition observed for drug-treated OCI-AML-2 cells. (Figure ?(Figure2D2D) Combination of CAL-101 and Vps34-IN-1 recapitulates the dual inhibitory effect of PI3KD/V-IN-01 We next investigated whether or not the combination of the selective PI3K inhibitor, CAL-101,.The slide was then stained with 1% eosin Y solution for 10-30sec with agitation. selective PI3K and Vps34 inhibitors. Interestingly, we observed FLT3-ITD AML cells are more sensitive to PI3KD/V-IN-01 than the FLT3 wt expressing cells. In AML cell inoculated xenograft mouse model, PI3KD/V-IN-01 exhibited dose-dependent anti-tumor growth efficacies. These results suggest that dual inhibition of PI3K and Vps34 might be a useful approach to improve the PI3K inhibitor’s anti-tumor efficacy. AG-120 and IP kinase assay demonstrated only modest inhibition of mTORC1 kinase by PI3KD/V-IN-01 (EC50 of 1700 nM), which is much less potent than activity displayed against PI3K and vps34 kinases. (Supplemental Figure 3) In HeLa cells, PI3KD/V-IN-01 effectively prevented LC3BII accumulation in the presence of EBSS (Earle’s balanced salt solution) and HCQ (hydrochloroquine) with an EC50 of 413 nM. (Figure ?(Figure1E)1E) In addition, an immunofluorescence experiment showed that PI3KD/V-IN-01 increased LC3B puncta in HeLa cells in a dose-dependent way, which was similar to the Vps34 specific inhibitor Vps34-IN-1, but not for the PI3K inhibitor CAL-101 or pan-PI3K inhibitor GDC-0941. (Figure ?(Figure1F1F and Supplemental Figure 2) This is not surprising as HeLa cells express Vps34 but not PI3K. In accordance with the established function of Vps34 in membrane trafficking [16, 17], the lysosome marker LAMP1 localization was affected by PI3KD/V-IN-01 and Vps34-IN-1, however not GDC-0941 or CAL-101. (Figure ?(Figure1F)1F) These biochemical and cellular data demonstrate that PI3KD/V-IN-01 was a highly potent and selective PI3K/Vps34 dual inhibitor. Open in a separate window Figure 1 Biochemical and pharmacological characterization of PI3KD/V-IN-01A. Chemical structure of PI3KD/V-IN-01. B. ADP-Glo? Biochemical IC50 determination of PI3KD/V-IN-01 against a panel of PI3K-related kinases. C. Determination of the inhibitory effect of PI3KD/V-IN-01 against class I PI3Ks in the cellular context. Specifically, PI3K in NIH-3T3 cells with PDGF-BB stimulation; PI3K in NIH-3T3 cells with LPA stimulation; PI3K in RAW264.7 cells with C5a stimulation; PI3K in Raji cells with anti-IgM stimulation. D. Selectivity profile of PI3KD/V-IN-01 in the DiscoveRx’s KinomeScan? platform. E. Effect of PI3KD/V-IN-01 on autophagy in HeLa cells using co-culture of EBSS and HCQ (25 M) and investigating LC3BII expression. F. Immuno-fluorescent imaging analysis of the effect of PI3K inhibitors on LC3BII expression in HeLa cells and of LAMP1 expression in HeLa cells treated with PI3K inhibitors. Table 1 Quantification of PI3KD/V-IN-01 EC50 against class I PI3Ks IP kinase assay. Activity of the downstream AKT kinase mediator, GSK3, and downstream transcription regulator, NFB, were not affected in any of the cell lines. What is noteworthy is that in MEC-2 cells, pERK was significantly inhibited by PI3KD/V-IN-01, GDC-0941 and CAL-101, but this phenomenon was not observed in OCI-AML-2, OCI-AML-3 and MV4-11 cells. These results demonstrate that PI3K is effectively inhibited in the cellular context, however differential responsiveness of certain signaling molecules to the inhibitors tested suggest that these cells might rely on different genetic/signaling network backgrounds. Open in a separate window Figure 2 Effect of PI3KD/V-IN-01 on cellular signaling, cell cycle progression, and autophagyA. Effect of PI3KD/V-IN-01 on PI3K- mediated signaling pathways in OCI-AML-2 (AML), MV4-11 (AML), OCI-AML-3(AML) and MEC-2 (CLL) cell lines. B. Apoptotic effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. C. Autophagy interruption effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. D. Cell cycle progression effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. We next examined the effect of PI3KD/V-IN-01 on induction of apoptosis. Interestingly, only for MV4-11 was apparent PARP cleavage observed, which clearly suggests drug-induced apoptosis of this line. (Figure ?(Figure2B)2B) For OCI-AML-2 and.Staining was developed with DAB; slides were counterstained with hematoxylin, dehydrated and mounted. HE staining HE staining was carried out as previously described.[15] First, the sections were hydrated and then the slide was dipped into a Coplin jar containing Mayer’s hematoxylin and agitated for 30 sec. known selective PI3K and Vps34 inhibitors. Interestingly, we observed FLT3-ITD AML cells are more sensitive to PI3KD/V-IN-01 than the FLT3 wt expressing cells. In AML cell inoculated xenograft mouse model, PI3KD/V-IN-01 exhibited dose-dependent anti-tumor growth efficacies. These results suggest that dual inhibition of PI3K and Vps34 might be a useful approach to improve the PI3K inhibitor’s anti-tumor effectiveness. and IP kinase assay shown only moderate inhibition of mTORC1 kinase by PI3KD/V-IN-01 (EC50 of 1700 nM), which is much less potent than activity displayed against PI3K and vps34 kinases. (Supplemental Number AG-120 3) In HeLa cells, PI3KD/V-IN-01 efficiently prevented LC3BII build up in the presence of EBSS (Earle’s balanced salt answer) and HCQ (hydrochloroquine) with an EC50 of 413 nM. (Number ?(Figure1E)1E) In addition, an immunofluorescence experiment showed that PI3KD/V-IN-01 increased LC3B puncta in HeLa cells inside a dose-dependent way, which was similar to the Vps34 specific inhibitor Vps34-IN-1, but not for the PI3K inhibitor CAL-101 or pan-PI3K inhibitor GDC-0941. (Number ?(Number1F1F and Supplemental Number 2) This is not amazing as HeLa cells express Vps34 but not PI3K. In accordance with the founded function of Vps34 in membrane trafficking [16, 17], the lysosome marker Light1 localization was affected by PI3KD/V-IN-01 and Vps34-IN-1, however not GDC-0941 or CAL-101. (Number ?(Figure1F)1F) These biochemical and cellular data demonstrate that PI3KD/V-IN-01 was a highly potent and selective PI3K/Vps34 dual inhibitor. Open in a separate window Number 1 Biochemical and pharmacological characterization of PI3KD/V-IN-01A. Chemical structure of PI3KD/V-IN-01. B. ADP-Glo? Biochemical IC50 dedication of PI3KD/V-IN-01 against a panel of PI3K-related kinases. C. Dedication of the inhibitory effect of PI3KD/V-IN-01 against class I PI3Ks in the cellular context. Specifically, PI3K in NIH-3T3 cells with PDGF-BB activation; PI3K in NIH-3T3 cells with LPA activation; PI3K in Natural264.7 cells with C5a activation; PI3K in Raji cells with anti-IgM activation. D. Selectivity profile of PI3KD/V-IN-01 in the DiscoveRx’s KinomeScan? platform. E. Effect of PI3KD/V-IN-01 on autophagy in HeLa cells using co-culture of EBSS and HCQ (25 M) and investigating LC3BII manifestation. F. Immuno-fluorescent imaging analysis of the effect of PI3K inhibitors on LC3BII manifestation in HeLa cells and of Light1 manifestation in HeLa cells treated with PI3K inhibitors. Table 1 Quantification of PI3KD/V-IN-01 EC50 against class I PI3Ks IP kinase assay. Activity of the downstream AKT kinase mediator, GSK3, and downstream transcription regulator, NFB, were not affected in any of the cell lines. What is noteworthy is definitely that in MEC-2 cells, pERK was significantly inhibited by PI3KD/V-IN-01, GDC-0941 and CAL-101, but this trend was not observed in OCI-AML-2, OCI-AML-3 and MV4-11 cells. These results demonstrate that PI3K is definitely efficiently inhibited in the cellular context, however differential responsiveness of particular signaling molecules to the inhibitors tested suggest that these cells might rely on different genetic/signaling network backgrounds. Open in a separate window Number 2 Effect of PI3KD/V-IN-01 on cellular signaling, cell cycle progression, and autophagyA. Effect of PI3KD/V-IN-01 on PI3K- mediated signaling pathways in OCI-AML-2 (AML), MV4-11 (AML), OCI-AML-3(AML) and MEC-2 (CLL) cell lines. B. Apoptotic effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. C. Autophagy interruption effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 CYSLTR2 and MEC-2 cells. D. Cell cycle progression effect of PI3KD/V-IN-01 in OCI-AML-2, MV4-11, OCI-AML-3 and MEC-2 cells. We next examined the effect of PI3KD/V-IN-01 on induction of apoptosis. Interestingly, only for MV4-11 was apparent PARP cleavage observed, which clearly suggests drug-induced apoptosis of this line. (Number ?(Figure2B)2B) For OCI-AML-2 and MEC-2, there was only a moderate induction.