Fig. 1.

MAP2K3 inhibited cell proliferation and invasion in ESCC in vitro and in vivo. (A) Expression of p‐MAP2K3 and MAP2K3 was detected by western blot in KYSE150 and KYSE520 cells after MAP2K3 transfection or knockout. (B) Cell growth was detected by CCK8 after MAP2K3 transfection or knockout in KYSE150 and KYSE520 cells. (C) Colony formation assay was performed after MAP2K3 transfection or knockout in KYSE150 and KYSE520 cells. (D) Flow cytometry analysis of cell apoptosis caused by MAP2K3 transfection or knockout in KYSE150 and KYSE520 cells. (E) Western blot assay was performed to detect apoptosis biomarkers, cleaved (cl‐) PARP, and caspase 3, after MAP2K3 transfection or knockout in KYSE150 and KYSE520 cells. (F) Cell invasion ability was detected by Transwell assay after MAP2K3 transfection or knockout in KYSE150 and KYSE520 cells. (G) Six weeks after KYSE520 MAP2K3‐KO and control cells were inoculated into the armpits of nude mice (n = 5 each group). Tumor volume and mouse weight were measured after injection of the indicated ESCC cells. (H) The tumor weight and size was measured in the indicated time after injection. (I) The representative photographs of immunohistochemistry staining of MAP2K3 and Ki67 in tissues from control or MAP2K3‐KO groups of mice (scale bar: 400 µm, 50 µm, respectively). Error bars represent the SD from at least three independent biological replicates. (*P < 0.05; **P < 0.01; ***P < 0.001 by Student’s t‐test)