Figure 6. Rapamycin inhibited nicardipine-induced sensitization of TMZ against GSCs. SU4 and SU5 cells were treated with TMZ (400 μM), nicardipine (20 μM), TMZ (400 μM) + nicardipine (20 μM), and TMZ (400 μM) + nicardipine (20 μM) +rapamycin (RPA) (50 nM), respectively, for 48 h. Then cells were stained with PI and Annexin V-FITC for apoptosis analysis (A–D). Western blot analysis of Bcl-2 and Bax in GSCs was performed after treating with TMZ (400 μM), nicardipine (20 μM), TMZ (400 μM) + nicardipine (20 μM), and TMZ (400 μM) + nicardipine (20 μM) +RPA (50 nM), respectively for 48 h (E–G). The survival analysis of GSCs bearing mice after chemotherapy, Kaplan-Meier curve indicated combined treatment with TMZ and nicardipine had positive survival benefit (H). * p < 0.05, ** p < 0.01, ***p<0.001.