Extended Data Fig. 4: PFKM levels increase in differentiated muscle populations.
From: PFKM governs metabolic shifts throughout skeletal muscle differentiation
a, PFKM single cell RNA-seq from tissue-wide human atlas across cell-types represented as a UMAP from 24 tissues (500,000 cells)35. b, Single-cell and single-nucleus RNA-seq data from the human skeletal muscle atlas represented as a UMAP from 40 major muscle populations13. c, RNA-seq data from the human skeletal muscle atlas13 represented as a dot plot. Relative expression levels of PFKM and myogenic target genes across muscle stem cells (MuSC), slow-twitch myofibres (type I) and fast-twitch myofibres (type II). Paired box 7 (PAX7), myogenin (MYOG), myoblast determination protein (MYOD), myosin heavy chain 1 (MYH1). d, Pseudotime trajectory analysis of combined muscle stem cells (MuSC), slow-twitch myofibres and fast-twitch myofibres for PFKM, muscle stem cell marker PAX7 and myofibre marker actin α1 (ACTA1). Each dot represents a single cell, with the color gradient indicating gene expression level. Cells are colored by cell cluster, pseudotime, and pseudotime branch state. e, mRNA expression of myogenic target genes from bulk RNA-seq37 in cultured human muscle cell line during a 12 day differentiation time course. Cyclin-dependent kinase 1 (CDK1), DNA binding protein inhibitor 3 (ID3), myogenin (MYOG), myosin heavy chain 3 (MYH3), myosin heavy chain 8 (MYH8), and titin (TTN), (n = 2 biological replicates). Statistical significance was determined by one-way ANOVA with post-hoc Dunnett’s analysis and data are shown as mean ± s.e.m. (CDK1: P=0.0009, P=0.0007, P=0.0002, P=0.0005, P=0.0010, P=0.0007. ID3: all values P<0.0001. MYOG: P<0.0001, P<0.0001, P<0.0001, P<0.0001, P=0.0004, P=0.0002. MYH3: P=0.0425. MYH8: P=0.0014, P=0.0013, P=0.0130, P=0.0074. TTN: P=0.0017, P<0.0001, P=0.0001, P<0.0001, P<0.0001. All ns P>0.05). f, Immunoblotting (IB) of PFKM in C2C12 muscle cells during a 3 day differentiation time course. Right, protein levels normalized to loading control (Actin), assessed by densitometry of n = 3 biological replicates. Statistical significance was determined by one-way ANOVA with post-hoc Dunnett’s analysis and data are shown as mean ± s.e.m. Exact P values are shown in graphs.
