Figure 2. Cell cycle and pro-apoptotic effect of NVP-AUY922 and 17-AAG in adult and pediatric GB cell lines.
The adult (U87MG) and the pediatric (SF188) GB lines were treated with 5xIC50 concentrations (determined by SRB assay) of either NVP-AUY922 or 17-AAG at time 0 and harvested at indicated time points. A- Cell cycle profile at representative time points determine by DNA content analysis using propidium iodine (PI) staining and flow cytometry, B- Sub-G1 population as a percentage of total events, as determined by flow cytometry. Values are mean±SD of at least 3 independent experiments. C- Immunoblotting analysis of G1/S (CDK4, cyclin D1), G2/M (CDC2) cell cycle transition proteins, PARP and caspase cleavage (■ full length and - cleaved fragments). GAPDH was used as loading control. Results are representative of at least 3 independent experiments. Densitometry data corresponding to the western blots in Figure 2C and repeat experiments which gave similar results are shown in Supplementary Table S2A (n=3; the standard error expressed as a percent of mean was calculated and found to have an overall mean value of 20.3%).