FIGURE 4.
Factors involved in m6A‐regulated expression of ZNF677. (A) Results based on TCGA and GEPIA database (http://gepia.cancer‐pku.cn/index) showed the expression level of IGF2BP2 in renal cell carcinoma (RCC) tumours and normal tissues. (B) IGF2BP2 RIP‐qPCR analysis of ZNF677 mRNA in control or Mettl3‐overexpressing OSRC cells. (C and D) Western blot and RT‐qPCR analysis of ZNF677 and IGF2BP2 protein (C) and mRNA (D) expression in OSRC cells transfected with control vector or IGF2BP2 construct. (E and F) OSRC cells were transfected with control vector, IGF2BP2 construct, pmirGLO‐ZNF677‐CDS‐WT (E) or pmirGLO‐ZNF677‐CDS‐MUT4 (F) for 24 h and then further treated with Act‐D for the indicated times. The mRNA of ZNF677 was checked by RT‐qPCR. (G) YTHDF1 RIP‐qPCR analysis of ZNF677 mRNA in control or Mettl3‐overexpressing OSRC cells. (H) Control or Mettl3‐overexpressing OSRC cells were transfected with control vector or YTHDF1 construct for 24 h, the expression of ZNF677 was checked by Western blot analysis. (I) OSRC cells were transfected with control vector, YTHDF1 construct, pmirGLO‐ZNF677‐CDS‐WT and pmirGLO‐ZNF677‐CDS‐MUT4 for 24 h, the expression of ZNF677 was checked by Western blot analysis. NS, not significant; *p < .05 or **p < .01 indicates a significant difference between the indicated groups