As shown in Figure ?Figure2A,2A, treatment of XL388 in 786-0 cells dose-dependently increased the activity of caspase-3 and caspase-9, but not caspase-8. Pharmacological or shRNA-mediated inhibition of MEK-ERK pathway sensitized XL388-induced cytotoxicity in RCC cells. and activity of XL388 in RCC cells. As demonstrated, 786-0 RCC cells, cultured in 10% FBS medium, were treated with XL388 at applied concentration. Trypan blue staining assay results demonstrated that XL388 dose-dependently induced 786-0 cell loss of life (Amount ?(Figure1A).1A). Further, XL388 also shown a time-dependent response in eliminating 786-0 cells (Amount ?(Figure1A).1A). Significant cell loss of life was notified 48 hours after XL388 (100-1000 nM) treatment (Amount ?(Figure1A).1A). The IC50s of XL388 had been 714.32 66.19 nM, 351.26 28.54 nM and 271.35 15.37 nM after 48, 72 and 96 hours treatment (Figure ?(Figure1A).1A). Cell Keeping track of Package-8 (CCK-8) cell viability assay leads to Amount ?Amount1B1B further demonstrated that XL388 was cytotoxic when put into the cultured 786-0 cells. XL388 once again shown a dose-dependent response in inhibiting 786-0 cells (Amount ?(Figure1B1B). Open up in another window Amount 1 XL388 inhibits RCC cell success and proliferationRCC cell lines (786-0 cells and A498 cells), the principal individual RCC cells (two lines, RCC1 and RCC2) or the HK-2 proximal tubule epithelial cells had been either left neglected (C, same for any statistics) or activated with listed focus of XL388, cells had been cultured in the conditional moderate for used period additional, cell success A., E and B. and proliferation D and C. had been examined with the assays talked about in the written text. For every assay, n=5. Data had been always portrayed as mean regular deviation (SD) (Same for any figures). Experiments within this amount had been repeated four situations, and similar outcomes had been attained. *< 0.05 vs. C group. The aftereffect of XL388 on 786-0 cell proliferation was examined following. BrdU incorporation assay leads to Amount ?Amount1C1C showed that XL388, at 100-1000 nM, reduced BrdU ELISA OD significantly, indicating the anti-proliferative activity with the chemical substance. Likewise, 100-1000 nM of XL388 also significantly decreased the amount of proliferative 786-0 colonies (Amount ?(Figure1D).1D). Hence, XL388 was anti-proliferation against 786-0 cells indeed. Next, we examined XL388's activity in various other RCC cells. As showed, treatment with XL388 (500 nM, 72 hours) generally reduced the viability of A498 RCC cells [3, 4] and two principal individual RCC cells (RCC1 and RCC2, Amount ?Amount1E).1E). Intriguingly, same XL388 treatment was non-cytotoxic towards the HK-2 proximal tubule epithelial cells [4, 25]. These total results show that XL388 inhibits survival and proliferation of individual RCC cells. XL388 activates apoptosis in RCC cells Following, the potential aftereffect of XL388 on RCC cell apoptosis was examined. As proven in Amount ?Amount2A,2A, treatment of XL388 in 786-0 cells dose-dependently increased the experience of caspase-3 and caspase-9, however, not caspase-8. The last mentioned is an signal of extrinsic apoptotic pathway activation [26]. On the other hand, the amount of cells with TUNEL-positive nuclei was considerably increased pursuing XL388 (100-1000 nM) treatment (Amount ?(Amount2B),2B), which also increased single-stranded DNA (ssDNA) apoptosis ELISA OD worth (Amount ?(Figure2C).2C). These results indicated that XL388 provoked apoptosis in 786-0 cells clearly. To research the function of apoptosis in XL388-induced cytotoxicity, many caspase inhibitors had been applied. Results demonstrated which the caspase-9 inhibitor z-LEHD-CHO, the caspase-3 inhibitor z-DEVD-CHO as well as the skillet caspase inhibitor z-VAD-CHO all generally inhibited XL388 (500 nM)-induced apoptosis activation (TUNEL assay, Amount ?Amount2D)2D) and subsequent 786-0 cell lethality (Amount ?(Amount2E,2E, tested with the CCK-8 viability decrease). To check XL388's influence on apoptosis in various other RCC cells, TUNEL staining assay was used. Results demonstrated that XL388 (500 nM) provoked significant apoptosis in A498 RCC cells and both lines of principal RCC cells (Amount ?(Figure2F).2F). However, there is no significant apoptosis activation in XL388-treated HK-2 epithelial cells (Amount ?(Figure2F).2F). Collectively, these total results show that XL388 provokes apoptosis in RCC cells. Open in another window Amount 2 XL388 activates apoptosis in RCC cells786-0 or A498 RCC cells, the principal individual RCC cells (RCC1 and RCC2) or the HK-2 cells had been stimulated with used focus of XL388, cells had been further cultured in the conditional moderate for applied period, cell apoptosis was examined with the caspase activity assay A., TUNEL staining assay F and B. as well as the ssDNA ELISA assay C. 786-0 cells had been pre-treated for 30 min with 50 M from the caspase-9 inhibitor z-LEHD-CHO (+lehd), the caspase-3 inhibitor z-DEVD-CHO (+devd) or the pan caspase inhibitor z-VAD-CHO (+vad), accompanied by XL388 (500 nM) treatment, cell viability and apoptosis were tested with the TUNEL assay D. as well as the CCK-8 assay E., respectively. For every assay, n=5. Tests in this amount had been repeated 3 x, and similar outcomes had been attained. *< 0.05 vs. C group. #< 0.05 vs. XL388 just group (D and E). XL388 blocks mTORC1 and mTORC2 in RCC cells We.Neurotox Res. loss of life (Amount ?(Figure1A).1A). Further, XL388 also shown a time-dependent response in eliminating 786-0 cells (Amount ?(Figure1A).1A). Significant cell loss of life was notified 48 hours after XL388 (100-1000 nM) treatment (Amount ?(Figure1A).1A). The IC50s of XL388 had been 714.32 66.19 nM, 351.26 28.54 nM and 271.35 15.37 nM after 48, 72 and 96 hours treatment (Figure ?(Figure1A).1A). Cell Keeping track of Package-8 (CCK-8) cell viability assay leads to Amount ?Amount1B1B further demonstrated that XL388 was cytotoxic when put into the cultured 786-0 cells. XL388 once again shown a dose-dependent response in inhibiting 786-0 cells (Amount ?(Figure1B1B). Open up in another window Amount 1 XL388 inhibits RCC cell success and proliferationRCC cell lines (786-0 cells and A498 cells), the principal individual RCC cells (two Indocyanine green lines, RCC1 and RCC2) or the HK-2 proximal tubule epithelial cells had been either left neglected (C, same for any statistics) or activated with listed focus of XL388, cells had been further cultured in the conditional medium for applied time, cell survival A., B and E. and proliferation C and D. were tested by the assays pointed out in the text. For each assay, n=5. Data were always expressed as mean standard deviation (SD) (Same for all those figures). Experiments in this physique were repeated four occasions, and similar results were obtained. *< 0.05 vs. C group. The potential effect of XL388 on 786-0 cell proliferation was tested next. BrdU incorporation assay results in Physique ?Physique1C1C showed that XL388, at 100-1000 nM, significantly decreased BrdU ELISA OD, indicating the anti-proliferative activity by the compound. Similarly, 100-1000 nM of XL388 also dramatically decreased the number of proliferative 786-0 colonies (Physique ?(Figure1D).1D). Thus, XL388 was indeed anti-proliferation against 786-0 cells. Next, we analyzed XL388's activity in other RCC cells. As exhibited, treatment with XL388 (500 nM, 72 hours) largely decreased the viability of A498 RCC cells [3, 4] and two main human RCC cells (RCC1 and RCC2, Physique ?Physique1E).1E). Intriguingly, same XL388 treatment was non-cytotoxic to the HK-2 proximal tubule epithelial cells [4, 25]. These results show that XL388 inhibits survival and proliferation of human RCC cells. XL388 activates apoptosis in RCC cells Next, the potential effect of XL388 on RCC cell apoptosis was tested. As shown in Physique ?Determine2A,2A, treatment of XL388 in 786-0 cells dose-dependently increased the activity of caspase-3 and caspase-9, but not caspase-8. The latter is an indication of extrinsic apoptotic pathway activation [26]. In the mean time, the number of cells with TUNEL-positive nuclei was significantly increased following XL388 (100-1000 nM) treatment (Physique ?(Physique2B),2B), which also increased single-stranded DNA (ssDNA) apoptosis ELISA OD value (Physique ?(Figure2C).2C). These results clearly indicated that XL388 provoked apoptosis in 786-0 cells. To investigate the function of apoptosis in XL388-induced cytotoxicity, several caspase inhibitors were applied. Results showed that this caspase-9 inhibitor z-LEHD-CHO, the caspase-3 inhibitor z-DEVD-CHO and the pan caspase inhibitor z-VAD-CHO all largely inhibited XL388 (500 nM)-induced apoptosis activation (TUNEL assay, Physique ?Physique2D)2D) and subsequent 786-0 cell lethality (Physique ?(Physique2E,2E, tested by the CCK-8 viability reduction). To test XL388's effect on apoptosis in other RCC cells, TUNEL staining assay was applied. Results showed that XL388 (500 nM) provoked significant apoptosis in A498 RCC cells and the two lines of main RCC cells (Physique ?(Figure2F).2F). Yet, there was no significant apoptosis activation in XL388-treated HK-2 epithelial cells (Physique ?(Figure2F).2F). Collectively, these results show that XL388 provokes apoptosis in RCC cells. Open in a separate window Physique 2 XL388 activates apoptosis in RCC cells786-0 or A498 RCC cells, the primary human RCC cells (RCC1 and RCC2) or the HK-2 cells were stimulated with applied concentration of XL388, cells were further cultured in the conditional medium for applied time, cell apoptosis was tested by the caspase activity assay A., TUNEL staining assay B and F. and the ssDNA ELISA assay C. 786-0 cells were pre-treated for 30 min with 50 M of the caspase-9 inhibitor z-LEHD-CHO (+lehd), the caspase-3 inhibitor z-DEVD-CHO (+devd) or the pan caspase inhibitor z-VAD-CHO (+vad), followed by XL388 (500 nM) treatment, cell apoptosis and viability were tested by the TUNEL assay D. and the CCK-8 assay E., respectively. For each assay, n=5. Experiments in this physique were.and activity of XL388 in RCC cells. As exhibited, 786-0 RCC cells, cultured in 10% FBS medium, were treated with XL388 at applied concentration. Trypan blue staining assay results exhibited that XL388 dose-dependently induced 786-0 cell death (Physique ?(Figure1A).1A). Further, XL388 also displayed a time-dependent response in killing 786-0 cells (Physique ?(Figure1A).1A). Significant cell death was notified 48 hours after XL388 (100-1000 nM) treatment (Physique ?(Figure1A).1A). The IC50s of XL388 were 714.32 66.19 nM, 351.26 28.54 nM and 271.35 15.37 nM after 48, 72 and 96 hours treatment (Figure ?(Figure1A).1A). Cell Counting Kit-8 (CCK-8) cell viability assay results in Physique ?Determine1B1B further demonstrated that XL388 was cytotoxic when added to the cultured 786-0 cells. XL388 again displayed a dose-dependent response in inhibiting 786-0 cells (Physique ?(Figure1B1B). Open in a separate window Physique 1 XL388 inhibits RCC cell survival and proliferationRCC cell lines (786-0 cells and A498 cells), the primary human RCC cells (two lines, RCC1 and RCC2) or the HK-2 proximal tubule epithelial cells were either left untreated (C, same for all those figures) or stimulated with listed concentration of XL388, cells were further cultured in the conditional medium for applied time, cell survival A., B and E. and proliferation C and D. were tested by the assays pointed out in the text. For each assay, n=5. Data were always expressed as mean standard deviation (SD) (Same for all those figures). Experiments in this physique were repeated four occasions, and similar results were obtained. *< 0.05 vs. C group. The potential aftereffect of XL388 on 786-0 cell proliferation was examined following. BrdU incorporation assay leads to Shape ?Shape1C1C showed that XL388, at 100-1000 nM, significantly reduced BrdU ELISA OD, indicating the anti-proliferative activity from the chemical substance. Likewise, 100-1000 nM of XL388 also significantly decreased the amount of proliferative 786-0 colonies (Shape ?(Figure1D).1D). Therefore, XL388 was certainly anti-proliferation against 786-0 cells. Next, we researched XL388's activity in additional RCC cells. As proven, treatment with XL388 (500 nM, 72 hours) mainly reduced the viability of A498 RCC cells [3, 4] and two major human being RCC cells (RCC1 and RCC2, Shape ?Shape1E).1E). Intriguingly, same XL388 treatment was non-cytotoxic towards the HK-2 proximal tubule epithelial cells [4, 25]. These outcomes display that XL388 inhibits success and proliferation of human being RCC cells. XL388 activates apoptosis in RCC cells Following, the potential aftereffect of XL388 on RCC cell apoptosis was examined. As demonstrated in Shape ?Shape2A,2A, treatment of XL388 in 786-0 cells dose-dependently increased the experience of caspase-3 and caspase-9, however, not caspase-8. The second option is an sign of extrinsic apoptotic pathway activation [26]. In the meantime, the amount of cells with TUNEL-positive nuclei was considerably increased pursuing XL388 (100-1000 nM) treatment (Shape ?(Shape2B),2B), which also increased single-stranded DNA (ssDNA) apoptosis ELISA OD worth (Shape ?(Figure2C).2C). These outcomes obviously indicated that XL388 provoked apoptosis in 786-0 cells. To research the function of apoptosis in XL388-induced cytotoxicity, many caspase inhibitors had been applied. Results demonstrated how the caspase-9 inhibitor z-LEHD-CHO, the caspase-3 inhibitor z-DEVD-CHO as well as the skillet caspase inhibitor z-VAD-CHO all mainly inhibited XL388 (500 nM)-induced apoptosis activation (TUNEL assay, Shape ?Shape2D)2D) and subsequent 786-0 cell lethality (Shape ?(Shape2E,2E, tested from the CCK-8 viability decrease). To check XL388's influence on apoptosis in additional RCC cells, TUNEL staining assay was used. Results demonstrated that XL388 (500 nM) provoked significant apoptosis in A498 RCC cells and both lines of major RCC cells (Shape ?(Figure2F).2F). However, there is no significant apoptosis activation in XL388-treated HK-2 epithelial cells (Shape ?(Figure2F).2F). Collectively, these outcomes display that XL388 provokes apoptosis in RCC cells. Indocyanine green Open up in another window Shape 2 XL388 activates apoptosis in RCC cells786-0 or A498 RCC cells, the principal human being RCC cells (RCC1 and RCC2) or the HK-2 cells had been stimulated with used focus of XL388, cells had been additional cultured in the conditional moderate for applied period, cell apoptosis was examined from the caspase activity assay A., TUNEL staining assay B and F. as well as the ssDNA ELISA assay C. 786-0 cells had been pre-treated for 30 min with 50 M from the caspase-9 inhibitor z-LEHD-CHO (+lehd), the caspase-3 inhibitor z-DEVD-CHO (+devd) or the pan caspase inhibitor z-VAD-CHO (+vad), accompanied by XL388 (500 nM) treatment, cell viability and apoptosis were tested from the.It has been proven that both mTOR complexes, mTORC2 and mTORC1, are important for most cancerous manners of RCC [6, 14]. used focus. Trypan blue staining assay outcomes proven that XL388 dose-dependently induced 786-0 cell loss of life (Shape ?(Figure1A).1A). Further, XL388 also shown a time-dependent response in eliminating 786-0 cells (Shape ?(Figure1A).1A). Significant cell loss of life was notified 48 Indocyanine green hours after XL388 (100-1000 nM) treatment (Shape ?(Figure1A).1A). The IC50s of XL388 had been 714.32 66.19 nM, 351.26 28.54 nM and 271.35 15.37 nM after 48, 72 and 96 hours treatment (Figure ?(Figure1A).1A). Cell Keeping track of Package-8 (CCK-8) cell viability assay leads to Shape ?Shape1B1B further demonstrated that XL388 was cytotoxic when put into the cultured 786-0 cells. XL388 once again shown a dose-dependent response in inhibiting 786-0 cells (Shape ?(Figure1B1B). Open up in another window Shape 1 XL388 inhibits RCC cell success and proliferationRCC cell lines (786-0 cells and A498 cells), the principal human being RCC cells (two lines, RCC1 and RCC2) or the HK-2 proximal tubule epithelial cells had been either left neglected (C, same for many numbers) or activated with listed focus of XL388, cells had been additional cultured in the conditional moderate for applied period, cell success A., B and E. and proliferation C and D. had been examined from the assays stated in the written text. For every assay, n=5. Data had been always indicated as mean regular deviation (SD) (Same for many figures). Experiments with this shape had been repeated four instances, and similar outcomes had been acquired. *< 0.05 vs. C group. The aftereffect of XL388 on 786-0 cell proliferation was examined following. BrdU incorporation assay leads to Shape ?Shape1C1C showed that XL388, at 100-1000 nM, significantly reduced BrdU ELISA OD, indicating the anti-proliferative activity from the chemical substance. Likewise, 100-1000 nM of XL388 also significantly decreased the amount of proliferative 786-0 colonies (Shape ?(Figure1D).1D). Therefore, XL388 was certainly anti-proliferation against 786-0 cells. Next, we researched XL388's activity in additional RCC cells. As proven, treatment with XL388 (500 nM, 72 hours) mainly reduced the viability of A498 RCC cells [3, 4] and two major human being RCC cells (RCC1 and RCC2, Shape ?Shape1E).1E). Intriguingly, same XL388 treatment was non-cytotoxic towards the HK-2 proximal tubule epithelial cells [4, 25]. These outcomes display that XL388 inhibits success and proliferation of human being RCC cells. XL388 activates apoptosis in RCC cells Following, the potential aftereffect of XL388 on RCC cell apoptosis was examined. As demonstrated in Shape ?Shape2A,2A, treatment of XL388 in 786-0 cells dose-dependently increased the experience of caspase-3 and caspase-9, however, not caspase-8. The second option is an sign of extrinsic apoptotic pathway activation [26]. In the meantime, the amount of cells with TUNEL-positive nuclei was considerably increased pursuing XL388 (100-1000 nM) treatment (Shape ?(Shape2B),2B), which also increased single-stranded DNA (ssDNA) apoptosis ELISA OD worth (Shape ?(Figure2C).2C). These outcomes obviously indicated that XL388 provoked apoptosis in 786-0 cells. To research the function of apoptosis in XL388-induced cytotoxicity, many caspase inhibitors had been applied. Results demonstrated how the caspase-9 inhibitor z-LEHD-CHO, the caspase-3 inhibitor z-DEVD-CHO as well as the Nog skillet caspase inhibitor z-VAD-CHO all mainly inhibited XL388 (500 nM)-induced apoptosis activation (TUNEL assay, Shape ?Shape2D)2D) and subsequent 786-0 cell lethality (Shape ?(Shape2E,2E, tested from the CCK-8 viability decrease). To check XL388’s influence on apoptosis in additional RCC cells, TUNEL staining assay was used. Results demonstrated that XL388 (500 nM) provoked significant apoptosis in A498 RCC cells and both lines of major RCC cells (Shape ?(Figure2F).2F). However, there is no significant apoptosis activation in XL388-treated HK-2 epithelial cells (Shape ?(Figure2F).2F). Collectively, these outcomes display that XL388 provokes apoptosis in RCC cells. Open up in another window Shape 2 XL388 activates apoptosis in RCC cells786-0 or A498 RCC cells, the principal human being RCC cells (RCC1 and RCC2) or the HK-2 cells had been stimulated with used focus of XL388, cells had been additional cultured in the conditional moderate for applied period, cell apoptosis was examined from the caspase activity assay A., TUNEL staining assay B and F. as well as the ssDNA ELISA assay C. 786-0 cells had been pre-treated for 30 min with 50 M from the caspase-9 inhibitor z-LEHD-CHO (+lehd), the caspase-3 inhibitor z-DEVD-CHO (+devd) or the pan caspase inhibitor z-VAD-CHO (+vad), accompanied by XL388 (500 nM) treatment, cell apoptosis and viability had been examined from the TUNEL assay D. as well as the CCK-8 assay E., respectively. For every assay, n=5. Tests in this shape had been repeated 3 x, and similar outcomes had been acquired. *< 0.05 vs. C group. #< 0.05 vs. XL388 just group (D and E). XL388 blocks mTORC1 and mTORC2 in RCC cells We following examined mTOR signaling in XL388-treated RCC cells. Treatment with XL388 (500 nM) in 786-0 RCC cells resulted in.Robinson CM, Ohh M. XL388 at used focus. Trypan blue staining assay outcomes proven that XL388 dose-dependently induced 786-0 cell loss of life (Shape ?(Figure1A).1A). Further, XL388 also shown a time-dependent response in eliminating 786-0 cells (Shape ?(Figure1A).1A). Significant cell loss of life was notified 48 hours after XL388 (100-1000 nM) treatment (Shape ?(Figure1A).1A). The IC50s of XL388 had been 714.32 66.19 nM, 351.26 28.54 nM and 271.35 15.37 nM after 48, 72 and 96 hours treatment (Figure ?(Figure1A).1A). Cell Keeping track of Package-8 (CCK-8) cell viability assay leads to Shape ?Shape1B1B further demonstrated that XL388 was cytotoxic when put into the cultured 786-0 cells. XL388 once again shown a dose-dependent response in inhibiting 786-0 cells (Shape ?(Figure1B1B). Open up in another window Shape 1 XL388 inhibits RCC cell success and proliferationRCC cell lines (786-0 cells and A498 cells), the principal human being RCC cells (two lines, RCC1 and RCC2) or the HK-2 proximal tubule epithelial cells had been either left neglected (C, same for many numbers) or activated with listed focus of XL388, cells had been additional cultured in the conditional moderate for applied period, cell success A., B and E. and proliferation C and D. had been examined from the assays described in the written text. For every assay, n=5. Data had been always indicated as mean regular deviation (SD) (Same for any figures). Experiments within this amount had been repeated four situations, and similar outcomes had been attained. *< 0.05 vs. C group. The aftereffect of XL388 on 786-0 cell proliferation was examined following. BrdU incorporation assay leads to Amount ?Amount1C1C showed that XL388, at 100-1000 nM, significantly reduced BrdU ELISA OD, indicating the anti-proliferative activity with the chemical substance. Likewise, 100-1000 nM of XL388 also significantly decreased the amount of proliferative 786-0 colonies (Amount ?(Figure1D).1D). Hence, XL388 was certainly anti-proliferation against 786-0 cells. Next, we examined XL388's activity in various other RCC cells. As showed, treatment with XL388 (500 nM, 72 hours) generally reduced the viability of A498 RCC cells [3, 4] and two principal individual RCC cells (RCC1 and RCC2, Amount ?Amount1E).1E). Intriguingly, same XL388 treatment was non-cytotoxic towards the HK-2 proximal tubule epithelial cells [4, 25]. These outcomes present that XL388 inhibits success and proliferation of individual RCC cells. XL388 activates apoptosis in RCC cells Following, the potential aftereffect of XL388 on RCC cell apoptosis was examined. As proven in Amount ?Amount2A,2A, treatment of XL388 in 786-0 cells dose-dependently increased the experience of caspase-3 and caspase-9, however, not caspase-8. The last mentioned is an signal of extrinsic apoptotic pathway activation [26]. On the other hand, the amount of cells with TUNEL-positive nuclei was considerably increased pursuing XL388 (100-1000 nM) treatment (Amount ?(Amount2B),2B), which also increased single-stranded DNA (ssDNA) apoptosis ELISA OD worth (Amount ?(Figure2C).2C). These outcomes obviously indicated that XL388 provoked apoptosis in 786-0 cells. To research the function of apoptosis in XL388-induced cytotoxicity, many caspase inhibitors had been applied. Results demonstrated which the caspase-9 inhibitor z-LEHD-CHO, the caspase-3 inhibitor z-DEVD-CHO as well as the skillet caspase inhibitor z-VAD-CHO all generally inhibited XL388 (500 nM)-induced apoptosis activation (TUNEL assay, Amount ?Amount2D)2D) and subsequent 786-0 cell lethality (Amount ?(Amount2E,2E, tested with the CCK-8 viability decrease). To check XL388's influence on apoptosis in various other RCC cells, TUNEL staining assay was used. Results demonstrated that XL388 (500 nM) provoked significant apoptosis in A498 RCC cells and both lines of principal RCC cells (Amount ?(Figure2F).2F). However, there is no significant apoptosis activation in XL388-treated HK-2 epithelial cells (Amount ?(Figure2F).2F). Collectively, these outcomes present that XL388 provokes apoptosis in RCC cells. Open up in another window Amount 2 XL388 activates apoptosis in RCC cells786-0 or A498 RCC cells, the principal individual RCC cells (RCC1 and RCC2) or the HK-2 cells had been stimulated with used focus of XL388, cells had been additional cultured in the conditional moderate for applied period, cell apoptosis was examined with the caspase activity assay A., TUNEL staining assay B and F. as well as the ssDNA ELISA assay C. 786-0 cells had been pre-treated for 30 min with 50 M from the caspase-9 inhibitor z-LEHD-CHO (+lehd), the caspase-3 inhibitor z-DEVD-CHO (+devd) or the pan caspase.