Fig. 2

SRT2183 induces glioma cell cycle arrest and apoptosis. a LN229 and U87MG cells were treated with vehicle or 10 μM SRT2183, the cells were stained with PI for cell cycle analysis after 24 h. Results portrayed as the mean ± SEM (***p < 0.001). and were analyzed by FACS after staining with propidium iodide for cell cycle analysis at 24 h. Results represent as the mean ± SEM (*p < 0.05). b LN229 and U87MG cells were treated with vehicle or 10 μM SRT2183 for 12, 24, 48 h, protein levels of CyclinD1, P-Rb, Rb, and GAPDH were analyzed by immunoblot (IB) (upper panel). LN229 and U87MG cells were treated with vehicle or 10 μM SRT2183 for 24 h. Location of ki67 identified by fluorescent immunostaining. Nucleus stained with DAPI (lower panel). c LN229 and U87MG cells were treated with vehicle or 10 μM SRT2183, the cells were assessed by immunofluorescence staining with ki67 and DAPI d LN229 and U87MG cells were treated with vehicle or 10 uM SRT2183 for 24 and 48 h. AnnexinV/7-AAD double-staining was using for apoptosis analysis by FACS, 5 uM concentration of Doxorubicin was taken as a positive control. e LN229 and U87MG cells were treated for 24 and 48 h using SRT2183 and expression of Caspase-3, PARP, Bim, Bcl-xl and Bcl-2 were examined by IB. 5 uM concentration of Doxorubicin were taken as a positive control. f LN229 and U87MG cells pre-treated with Z-VAD-FMK (50 uM) then with the vehicle control or 10 μM SRT2183 after 24 and 48 h, the number of viable cells were quantified by CCK-8 assay. The above experiments were performed three times, (*0.01 < p < 0.05, **0.001 < p < 0.01, ***p < 0.001, n.s. = not significant).