Fig. 2

ZA induces TSC2-null cell apoptosis by GGPP. a MTT assay of TSC2-null cells treated with control, as well as 25, 50, and 100 μM ZA. b Western blot analysis after treatment with ZA (25 and 50 μM, respectively; and 20 nM RAPA for 24 h) in TSC2-null cells. c Immunostaining of Ki67 after treatment with ZA (25 and 50 μM; and 20 nM RAPA for 24 h). Immunofluorescence staining was performed and analyzed in independent 3 experiments. d Apoptosis of TSC2-null cells treated with ZA assayed using the DeadEnd™ Fluorometric TUNEL System. The green dots represented apoptotic cells, and DAPI (blue) indicated cell nuclei. Scar bar, 100 and 200 μm, respectively. e Immunoblotting analysis of apoptosis marker (cleaved-caspase3) after treatment with ZA alone, the combination of ZA and FPP, and the combination of ZA and GGPP, respectively, for 24 h. The experiment was performed for three times. All data were presented as mean ± SEM. Compared with the control group, ** P < 0.01, *** P < 0.001