Fig. 3: Identification of Gys2 mRNA by global N6-methyladenosine modification analysis as a key substrate of METTL3 in mouse liver. | Nature Communications

Fig. 3: Identification of Gys2 mRNA by global N6-methyladenosine modification analysis as a key substrate of METTL3 in mouse liver.

From: N6-methyladenosine modification governs liver glycogenesis by stabilizing the glycogen synthase 2 mRNA

Fig. 3

a The most enriched sequence motif of m6A peaks in mRNAs from 8-week-old WT or cKO mouse hepatocytes (HC) or liver (LI) samples. b Distribution of m6A peaks along the 5′UTR, CDS, and 3′UTR regions of total mRNAs from 8-week-old mouse samples after normalized with length. c RNA-Seq and MeRIP-Seq identified downregulated and m6A-lost genes in Mettl3-cKO liver tissue or hepatocytes comparing to wild-type samples. All the samples were from 8-week-old mice. MeRIP-seq data were from hepatocytes of male mice; RNA-seq data were from hepatocytes and liver tissues of male mice and liver tissues of female mice, respectively. d m6A MeRIP-Seq revealed the location of specific m6A peak in Gys2 locus in hepatocytes of wildtype or Mettl3-cKO mice. //, deletion of a long and uninformative intron for better exhibition. e m6A enrichment of Gys2 mRNA in mettl3-HET or cKO hepatocytes by m6A-RIP-qPCR. n = 3 independent experiments. Data are presented as mean values +/− SEM. Two-way ANOVA was performed to determine a difference among each two groups. **p < 0.01 comparing with IgG:HET; ##p < 0.01 comparing with m6A:HET. f qRT-PCR of Gys2 mRNA levels (β-actin as reference) in 8-week-old mouse livers with indicated Mettl3 genotypes. n = 4 animals. Data are presented as mean values +/− SEM. Two-sided Student’s t test was performed to determine a difference among groups. **p = 0.004. g Western blotting assay of GYS2 protein levels in 8-week-old mouse livers with indicated Mettl3 genotypes. The ratio of GYS2 to β-actin were shown between two lanes. This experiment was repeated independently with similar results at least 3 times. ♂, male; ♀, female. h qRT-PCR of Gys2 mRNA levels (β-actin as reference) in wildtype mouse livers with indicated ages. Two-sided Student’s t test was performed to determine a difference among groups. **p = 0.004. n = 4 animals. Data are presented as mean values +/− SEM. Source data are provided as a Source data file.

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