Fig. 2 | Scientific Reports

Fig. 2

From: LMNA R482L mutation causes impairments in C2C12 myoblasts subpopulations, alterations in metabolic reprogramming during differentiation, and oxidative stress

Fig. 2

scRNA-seq of myoblasts C2C12 bearing hLMNA gene WT or R482L. (a) UMAP of 9470 hLMNA-WT/R482L C2C12 cells divided into 4 clusters and 2 populations MES and MYO. (b) Proportion of cells in clusters for WT and R482L myoblasts. (c) UMAP of cell cycle phase scoring. (d) Distribution of cell cycle phases in clusters for hLMNA-WT/R482L C2C12 myoblasts. (e) UMAP of cells’ pseudotime assessed with Monocle3. (f) Heatmap of the top 20 differentially expressed (DE) marker genes for each cell cluster in hLMNA-WT/R482L C2C12 myoblasts with log2FoldChange > 0.25, FDR = 0.01. (g) UMAP feature plots of the expression for some DE marker genes. (h) Immunocytological staining of WT and R482L myoblasts confirmed that all cells co-express Desmin and aSMA (Acta2). Scale bars represent 50 μm. (i) Western blotting (upper panel) analysis showed the upregulation of aSMA in R482L samples; fold changes (lower panel) of the indicated proteins relative to the WT protein expression were quantified by densitometric scanning; n = 3; ns—nonsignificant. Mann Whitney test was used. Uncropped blots images are in Fig. S2. (j,k) Pathways enriched in clusters based on DE marker genes and analyzed with Gene Ontology (GO) Biological Processes (h) and GO Molecular Functions (i) databases (FDR = 0.01).

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