Fig. 4
Characterization of m6A‒modified genes that are regulated by ALKBH5 in human epidermis samples and keratinocyte cell lines. a Flowchart of m6A modification downstream analysis. MeRIP-seq identified 3710 genes shared by human epidermis samples and keratinocyte cell lines, with specific m6A peaks in the 3’UTR. RNA-seq identified 306 downregulated genes after ALKBH5 knockdown; the cutoffs used to define downregulated genes were |FC|< 0.5 and P < 0.05. FC fold change. Finally, 16 candidate genes with both m6A peaks and differential expression were identified as potential downstream targets of ALKBH5. b GO enrichment map of 3710 genes. c The regulatory network of PELI2 and 15 other candidate genes generated by IPA. Please note that C3orf33, PGBD2, ZNF20, ZNF573, ZNF577, and ZNF785 are not shown as they have no pathway connections with PELI2. d The expression of 16 candidate genes after ALKBH5 knockdown was measured using qRT‒PCR. The experiments were performed in triplicate, and the relative mRNA expression is shown as the mean ± SD. One-way ANOVA, *P < 0.05; **P < 0.01. e IGV tracks displaying the MeRIP-seq read coverage of PELI2 in human epidermis and HaCaT. f m6A-RIP-qPCR assays confirmed the m6A modification of PELI2 transcripts. The experiments were performed in triplicate. The relative mRNA expression in the anti-m6A antibody group was compared to that in the IgG group, and it is shown as the mean ± SD. One-way ANOVA, **P < 0.01