Fig. 5: METTL3 is essential for m6A modification of Prdm16, Pparg, and Ucp1 transcripts.
From: METTL3 is essential for postnatal development of brown adipose tissue and energy expenditure in mice
a–c The read density from m6A-RIP-seq experiments on BKO–flox/flox pairs showing the m6A peaks identified in the Prdm16, Pparg, and Ucp1 transcripts. d–f The m6A modification in Prdm16, Pparg, and Ucp1 transcripts in iBAT of 8-week-old Mettl3flox/flox and BKO mice were measured by m6ARIP-RT-qPCR (n = 3 for each group). g–j Primary brown preadipocytes seeded in 24-well plates were co-transfected with pMIR-REPORT Luciferase vectors (Prdm16, Pparg, Ucp1), siRNAs (Scramble siRNA, siMettl3-1, siMettl3-2) and β-galactosidase (β-Gal) reporter plasmid by X-tremeGENE siRNA Transfection Reagent for 24 h. Cells were then induced to differentiate for 48 h. METTL3 and GAPDH protein levels were measured by immunoblotting g. Relative Prdm16, Pparg, and Ucp1 luciferase activity were measured and normalized to β-Gal levels h–j (Scr in h, n = 4; others, n = 5 biologically independent cell samples for each group). Data represent the mean ± SEM. Significance was determined by unpaired two-tailed Student’s t test analysis. *p < 0.05. **p < 0.01. Source data are provided as a Source Data file.
