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. 2022 Aug 17;25:101479. doi: 10.1016/j.tranon.2022.101479

Fig. 5.

Fig 5

miR-198 increases CXXC4 expression and inactivates the Wnt/β-catenin pathway by targeting EZH2. A, mRNA expression of CXXC4 in GAC tissues and adjacent non-neoplastic tissues determined by RT-qPCR. * p < 0.05, compared with adjacent non-neoplastic tissues. B, Immunohistochemical analysis of CXXC4 protein in GAC tissues and adjacent non-neoplastic tissues. C, mRNA expression of CXXC4 in GAC cells (GC9811-P, XGC-9811, AGS), and normal gastric mucosal cells (GES1) determined by RT-qPCR. * p < 0.05, compared with GES1 cells. D, Western blot analysis of CXXC4 and β-catenin proteins in GAC cells (GC9811-P, XGC-9811, AGS), and normal gastric mucosal cells (GES1). * p < 0.05, compared with GES1 cells. E, Enrichment of EZH2 in the promoter region of CXXC4 gene detected by ChIP. * p < 0.05, compared with IgG antibody. F, mRNA expression of EZH2 and CXXC4 in GC9811-P cells treated with si-EZH2 or combined with si-CXXC4 determined by RT-qPCR. * p < 0.05, compared with si-NC-treated GC9811-P cells. # p < 0.05, compared with si-EZH2-treated GC9811-P cells. G, Western blot analysis of expression of EZH2, CXXC4 and β-catenin proteins along with p-GSK-3β extent in GC9811-P cells treated with si-EZH2 or combined with si-CXXC4. * p < 0.05, compared with si-NC-treated GC9811-P cells. # p < 0.05, compared with si-EZH2-treated GC9811-P cells. H, miR-198 expression and mRNA expression of EZH2 and CXXC4 in GC9811-P cells treated with miR-198 mimic or combined with oe-EZH2 determined by RT-qPCR. * p < 0.05, compared with oe-NC + mimic NC-treated GC9811-P cells. * p < 0.05, compared with oe-NC + mimic NC-treated GC9811-P cells. # p < 0.05, compared with oe-NC + miR-198 mimic-treated GC9811-P cells. I, Western blot analysis of expression of EZH2, CXXC4 and β-catenin proteins along with p-GSK-3β extent in GC9811-P cells treated with miR-198 mimic or combined with oe-EZH2. * p < 0.05, compared with GC9811-P cells treated with oe-NC + mimic NC. # p < 0.05, compared with GC9811-P cells treated with oe-NC + miR-198 mimic. The cell experiment was conducted three times independently.