Fig. 1: YTHDF2 regulates postnatal skeletal muscle mass.
From: YTHDF2 governs muscle size through a targeted modulation of proteostasis
a Schematic of muscle-specific YTHDF2 knockout (Y2-KO) mouse generation, created in Adobe Illustrator. b qPCR analysis of Ythdf2 mRNA expression using the gastrocnemius of control (Ctrl) and Y2-KO mice. c Western blot analysis of YTHDF2 protein expression in Ctrl and Y2-KO gastrocnemius. d Quantification of YTHDF2 protein expression using total protein detection by Ponceau stain as a loading control. e Tibialis anterior weight, f quadriceps weight, and g gastrocnemius weight for 1- and 2-month-old Ctrl and Y2-KO mice, normalized to tibia length. h Body weight and i tibia length for Ctrl and Y2-KO mice at 2 months of age. j Representative hematoxylin/eosin (H&E) staining for Ctrl and Y2-KO tibialis anterior. k Quantification of m6A relative to total adenosine (m6A/A) as determined by ELISA in Ctrl and Y2-KO quadriceps. l Western blot analysis of YTHDF2 protein expression in Ctrl tibialis anterior compared to Y2-KO injected with AAV-GFP or AAV-YTHDF2 (AAV-Y2) at 2 months of age and analyzed 8 weeks following administration. m Tibialis anterior weight normalized to tibia length for the indicated groups. Biological animal replicates: n = 5 (Ctrl) and 4 (Y2-KO) in panel b; n = 3 (Ctrl) and 3 (Y2-KO) in panel c and d; n = 4 (Ctrl at 1 month), 5 (Y2-KO at 1 month), 12 (Ctrl at 2 months), and 8 (Y2-KO at 2 months) for panel e–g; n = 12 (Ctrl) and 8 (Y2-KO) in panel h, I; n = 5 (Ctrl) and 4 (Y2-KO) in panel k; n = 3 (Ctrl), 3 (Y2-KO + AAV-GFP), and 3 (Y2-KO + AAV-Y2) in panel l; n = 9 (Ctrl), 6 (Y2-KO + AAV-GFP), and 7 (Y2-KO + AAV-Y2) in panel m. Data were presented as the mean ± SEM with the individual biological samples shown. Significance was determined by two-tailed Student’s t-test for comparisons between Ctrl and Y2-KO mice, by two-way ANOVA with Tukey’s HSD multiple-comparison test for comparison of the means of Ctrl and Y2-KO mice at 1 and 2 months of age, or by one-way ANOVA for Ctrl, Y2-KO + AAV-GFP, and Y2-KO + AAV-Y2 mice: *p ≤ 0.05, **p ≤ 0.01, ****p ≤ 0.0001. Scale bar = 100 µm for panel j.
