Figure 6.

ZC3H13-mediated m⁶A modification reduces PKM2 mRNA stability in HCC cells. (a, b) Detection of the remaining PKM2 transcript in Hep3B as well as HUH-7 cells with ZC3H13 knockdown following exposure to Act D for the indicated time points. (c, d) Detection of the remaining PKM2 transcript for Hep3B as well as HUH-7 cells with ZC3H13 overexpression under exposure to Act D for the indicated time points. (e) RIP assay examining the enrichment levels of PKM2 mRNA in Hep3B as well as HUH-7 cells under incubation by anti-ZC3H13 or anti-IgG antibody. GAPDH transcript was utilized as a control. (f, g) Luciferase reporter assay examining the effects of ZC3H13 on wild-type PKM2 (PKM2-WT) or mutant PKM2 (PKM2-MUT) vector for Hep3B as well as HUH-7 cells. (h) MeRIP‐qPCR detecting m⁶A modification levels of PKM2 through immunoprecipitation of m⁶A-modified mRNA for Hep3B as well as HUH-7 cells with empty vector or ZC3H13 overexpression. Ns: not significant; ^∗P < 0.05; ^∗∗P < 0.01; ^∗∗∗P < 0.001; and ^∗∗∗∗P < 0.0001.