Research Paper Volume 12, Issue 18 pp 18384—18395

Figure 5. Ectopic overexpression of p38γ promotes human OS cell progression in vitro. Expression of listed genes in the stable OS1 cells, with the pLenti6-puro-GFP-p38γ expression vector (p38γ-OE-sL1 and p38γ-OE-sL2, two lines) or the empty vector (“Vec”), tested by Western blotting and qPCR assays (A–C); Cells were further cultured for applied time periods, cell growth (cell counting assay, D), proliferation (by measuring EdU ratio, E) and migration (“Transwell” assay, F) were tested; Rb phosphorylation and cyclin E1/A expression were tested by Western blotting (G). Rb phosphorylation and cyclin E1/A in the OS1 cells with scramble control shRNA (“shC”) or the applied p38γ shRNA (p38γ-shRNA-s1/s2), as well as in the p38γ-KO OS1 cells (by sgRNA-1), were tested and results were shown (H). Expression of listed proteins was quantified and normalized to the loading control (B, G, H). Data presented as mean ± standard deviation (SD, n=5). ** p< 0.01 vs. “Vec” cells.*** p< 0.001 vs. “Vec” cells. Experiments in this figure were repeated five times.