Fig. 3

SAHA induces apoptosis in GSCs. a and b GSCs from U87MG cells were treated with different doses of SAHA as indicated for 2 days. After treatment, cells were stained with PI alone (a) and with both FITC Annexin V and PI (b), and apoptotic cell death was measured using flow cytometry. c and d Percentage of sub-G1, early and late apoptotic cells. Data are presented as mean ± S.D. for experiments performed in triplicate; bar, S.D. e Expression of cleaved-caspase-8, −caspase-3, and -PARP as well as pro-caspase-9 were determined by Western blotting. f and g GSCSs were treated with 25 μM SAHA alone, 20 μM Z-VAD-FMK (Caspase Inhibitor) alone, or SAHA + Z-VAD-FMK for 48 h, following which cell lysates were analyzed via Western blot with indicated antibodies (f). g Cell viability was also assessed using MTT assay. Bar graphs represent means ± SEM from three independent experiments (t-test: **p < 0.01)