Fig. S5.

MAP2K4 suppresses malignant phenotypes caused by miR-BART22 in NPC cells.

(a)(b) QPCR(n = 3 independent experiments, Student's t-test) and Western blot was used to examine the MAP2K4 mRNA and protein expressions in miR-BART22-overexpressed NPC cells and their control cells. (c) Tumor sphere formation experiment generated by the miR-BART22 NPC cells transfected with MAP2K4 and their empty control cells are shown; three independent experiments were performed. Original magnification, ×100, scale bar, 200 μm; (d) Transwell and Boyden Chamber assays, (e) Dose-response curves of HONE1-EBV⁺ and 5-8F treated with miR-BART22 inhibitor or controls. Experiments were repeated three times with similar results, mean ± SD, *P < .05. (f) Western blot analysis of PI3K, p-PI3K, AKT, p-AKT, c-Jun, c-Myc, GSK3β, p-GSK3β, β-catenin, E-cadherin, N-cadherin, Vimentin, Nanog, OCT4, Sox2, Ubc and TRAF6 expression in BART22-overexpressed NPC cells with MAP2K4 transfection. β-actin served as controls. (g) Co-immunoprecipitation analysis of ubiquitin in BART22 overexpressed HONE1-EBV⁺ cells with MAP2K4 transfected. (h–i) Chromatin immunoprecipitation analysis of c-Jun binding to the transcriptional regulatory region of MYH9 in BART22 overexpressed HONE1-EBV⁺ and 5-8F cells with MAP2K4 transfected. All data are presented as the mean ± SD. Experiments were repeated three times. (j)(k) QPCR was used to examine the Ubc and MYH9 mRNA levels in BART22 overexpressed NPC cells with MAP2K4 transfected (n = 3 independent experiments, Student's t-test).