Extended Data Fig. 3: Loss of the m⁶A methylase METTL14 increases lipogenesis and hepatic triglyceride accumulation.

a, Western blot of Mettl14 in other tissues from WT and Mettl14 L-KO chow-fed males. Equal amounts of protein were pooled from four animals per group and run in triplicate. b, Lipidomics heatmap comparing hepatic lipidome of WT versus Mettl14 L-KO chow-fed males. (n = 8 per group). c, Quantification of various lipid species from lipidomics analysis of chow-fed male livers (n = 8). Statistical analysis was performed using unpaired two-tailed t-test. Values are mean ± SEM. d, Quantification of total food intake for chow-fed male WT and Mettl14 L-KO mice (n = 8 per group). e, Body weight and percent fat measured by MRI for chow-fed male WT and Mettl14 L-KO mice (n = 8 WT mice, n = 7 L-KO mice). f, Quantification of cellular respiratory rate in livers of chow-fed WT and Mettl14 L-KO mice using NADH as the acceptor (n = 5 per group). g, qPCR analysis of fatty acid oxidation gene expression from chow-fed WT and Mettl14 L-KO livers (n = 5). The experiment was repeated two times with similar results. h, Quantification of serum triglyceride levels from chow-fed male WT and Mettl14 L-KO mice (n = 8 per group). i, qPCR analysis of gene expression from liver of WT and Mettl14 L-KO mice (n = 4 per group). The experiment was repeated two times with similar results. j, Western blot of lipogenic protein levels in a second independent cohort of chow-fed male mice. Equal amounts of protein were pooled from eight animals per group and run in triplicate. The experiment was repeated three times with similar results. k, Western blot comparing levels of lipogenic proteins in the setting of chronic Mettl14 deficiency (albumin-cre transgenics or control). Equal amounts of protein were pooled from five animals per group and run in triplicate. The experiment was repeated two times with similar results. l, Quantification of hepatic triglyceride concentrations from livers of albumin-cre transgenic or control mice (n = 4 Cre-N mice, n = 5 Cre-P mice). m, Quantification of lipogenic transcript m⁶A abundance in WT and Mettl14 L-KO chow-fed livers measured by m⁶A-IP-qPCR analysis (n = 4). The relative enrichment of m⁶A in each sample was determined by normalizing to ten-fold input. The experiment was repeated two times with similar results. n, Quantification of lipogenic transcript m⁶A abundance in WT and Mettl14 L-KO chow-fed livers measured by m⁶A-IP-qPCR analysis (n = 4). The relative enrichment of m⁶A in each sample was determined by normalizing to GAPDH. The experiment was repeated two times with similar results. Mice were fed chow diet for 4 weeks and fasted for 4-hrs. prior to sacrifice. Values are mean ± s.e.m. of 4 (I,l-n), 5 (f,g), 6 (d), or 8 (c,e,h) independent biological replicates. P values were calculated using unpaired two-tailed t-test. (c-i,l-n). *P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. The precise n, P values, and details of the statistical testing are provided in the source data file.

Source data