Fig. 4
(A) The four-quadrant plots showed significant changes in mRNA expression and m6A methylation in gene distribution. (B) The mRNA level of Appl1, Ntrk2, Trub2, Cbx4 and Tlr4 (n = 3 or 4). (C) Dot blot detecting the global m6A modification of mRNAs. Methylene blue (MB) staining was used as a loading control. Gel images were cropped, and the original gels were presented in Supplementary Figure S2. (D) MeRIP-seq detected the m6A abundance of Trub2 mRNA in the HFD group and HB group. The yellow highlight showed that the intensity of m6A peak was significantly higher in the HB group than in the HFD group. (E) The protein level of Trub2 in the liver of CK, HFD, HB mice (n = 3). Gel images were cropped, and the original gels were presented in Supplementary Figure S3. (F) Quantitative results of Trub2 protein levels. (G-H) Western blot analysis was employed to assess and quantify the expression efficiency of Trub2 in OA/PA-treated AML12 cells after its overexpression (n = 4). Gel images were cropped, and the original gels were presented in Supplementary Figure S4. (I) Triglyceride levels in AML12 cells after transfected with Trub2 after OA/PA treatment (n = 3). (J) Triglyceride content in AML12 cells after transfected with siNC and siTrub2 (n = 6). The data were expressed as the mean ± SD. *P < 0.05, **P < 0.01, ****P < 0.0001. CK, Control mice; HFD, High fat diet; HB, High fat diet with betaine.
