Fig. 4.

EGFR depletion recapitulates the inhibitory effect of miR-1231 on glioma cell proliferation. a Immunohistochemical analysis of EGFR and Ki-67 in tumors derived from negative control (si-NC) and EGFR knockdown (si-EGFR) cells in vivo. Scale bars, 100 µm. b Western blot analysis of the indicated proteins in cells transfected with siRNA negative control or siRNA targeting EGFR. c CCK-8 assay of the proliferation rate of cells after transfection with si-NC or si-EGFR following culture for 96 h. Data are presented as the means of triplicate experiments, *P < 0.05, **P < 0.01. d, e Colony formation assays were performed in LN229, U251, and PG1 cell lines stably expressing si-NC or si-EGFR. Data are presented as the means of triplicate experiments, **P < 0.01. f, g EDU assays in LN229, U251, and PG1 cell lines stably expressing si-NC or si-EGFR 48 h after transfection. Data are presented as the means of triplicate experiments, **P < 0.01. h, i Flow cytometry analysis and cell cycle distribution in LN299, U251, and PG1 cells transfected with si-EGFR or si-NC for 72 h