Figure 6.

m⁶A-modified LEAWBIH activated Wnt/β-catenin signaling. (A) GSEA of HCC subclass S1 and Wnt/β-catenin signaling gene signatures in LEAWBIH high expression group versus LEAWBIH low expression group. NES, normalized enrichment score. (B and C) β-catenin reporter TOPFlash was co-transfected with pRL-TK into HuH-7 cells with overexpression of wild-type or mutated LEAWBIH (B) or HuH-7 cells with knockdown of LEAWBIH (C). Luciferase activities were measured 48 h after transfection. Results are presented as the relative ratio of firefly luciferase activity to Renilla luciferase activity. (D) The expression of Wnt/β-catenin targets in HuH-7 cells with overexpression of wild-type or mutated LEAWBIH was measured by qPCR. (E) The expression of Wnt/β-catenin signaling targets in HuH-7 cells with knockdown of LEAWBIH was measured by qPCR. For (B–E), results are presented as mean ± SD of 3 independent experiments. *P < 0.05, **P < 0.01, ns, not significant, by one-way ANOVA followed by Dunnett’s multiple comparisons test. (F–I) The correlation between LEAWBIH expression and Wnt/β-catenin signaling targets AXIN2 (F), LEF1 (G), CCND1 (H), or MYC (I) expression in 371 HCC tissues, based on the TCGA-LIHC RNA-seq data. r and P values were calculated by Spearman correlation analysis.