Fig. 4

(A) Effect of THPS and BAPTA on ANG II-induced mesangial cell apoptosis. Equal numbers of cells were incubated in media containing either buffer (control), ANG II (10–8 M), THPS (50 µM), BAPTA (50 µM), ANG II + THPS, or ANG II + BAPTA for 24 hr. Subsequently, cells were assayed for apoptosis. Results (means ± SEM) are from four series of experiments, each carried out in triplicate. *p < 0.001 compared with control, THPS, BAPTA, ANG II + BAPTA, ANG II + THPS; **p < 0.001 compared with control, ANG II and Ang II + THPS; ***p < 0.001 compared with ANG II and THPS alone. (B) Dose-response effect of thapsigargin (THPS) on mesangial cell apoptosis. Equal numbers of MMCs were incubated in media containing either vehicle or variable concentrations of THPS (1, 10, 100 nM) for 24 hr. Subsequently, MMCs were stained with H-33342 and PI. Results (means ± SEM) are from four series of experiments, each carried out in triplicate. *p < 0.01 compared with control; **p < 0.001 compared with control and THPS, 1 nM; ***p < 0.001 compared with control and THPS, 1–10 nM. (C) Effect of THPS and BAPTA on TGF-β-induced mesangial cell apoptosis. Equal numbers of MMCs were incubated in media containing buffer (control), TGF-β (1 nglml), THPS (10 nM), BAPTA (50 µM), TGF-β +BAPTA, or TGF-β + THPS for 24 hr. At the end of the incubation period, cells were assayed for apoptosis. Results (means ± SEM) are from four series of experiments, each carried out in triplicate. *p < 0.001 compared with control, THPS, BAPTA, TGF-b + BAPTA; **p < 0.001 compared with control; ***p < 0.05 compared with control. (D) Effect of PD-123319 and losartan on ANG II-induced mesangial cell apoptosis. Equal numbers of MMCs were incubated in media containing either buffer (control) or ANG II (10⁻⁸ M) in the absence or presence of PD-123319 (PD) or losaratan (LOS) for 24 hr. Cells were then stained for apoptosis. Results (means ± SEM) are from four series of experiments, each carried out in triplicate. *p < 0.001 compared with control, ANG II + PD and Ang II + LOS; **p < 0.01 compared with control; ***p < 0.05 compared with respective PD and LOS alone.