Fig. 3
p110α inhibitors decrease cell proliferation of neuroblastoma cells. a–d Colony formation analysis of NBL-S cells (a, b) and KELLY cells (c, d) upon the treatment of different PI3K inhibitors (1 µM). Pictures were captured after 12-day treatment and afterwards crystal violet staining (a, c). Quantifications of all pictures were conducted by Image J (b, d). e Cell viability showing the cell growth after siRNA transfection. KELLY cells were seeded in 96-well plates at 6 h after siPIK3CA (50 nM) or siControl (50 nM) transfection. Cell viability was then determined by MTT assay at different time points. Data were expressed as mean ± SEM. f Cell viability showing the cell growth after p110α overexpression in IMR-32 cells. Cells were transfected with vector control or p110α-Myc and re-seeded into 96-wells after 24 h. Cell viability was determined by MTT assay at different time points. Data were expressed as mean ± SEM. g, h cell cycle distribution of NBL-S cells and KELLY cells treated with PI3K inhibitors as indicated (1 µM). Neuroblastoma cells were fixed after 24-h treatment with the PI3K inhibitors. The DNA content was then determined by PI staining and flow cytometry analysis. Cell cycle distribution was defined with Flow Jo. Data from 3 repeats were summarized. Bar charts were presented as mean ± SEM. The Student’s t-test was used for the statistical analysis of the G0/G1 proportion between the inhibitor-treated groups and the vehicle group. *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant