Fig. 8.

Characterization of the effect of Myod1 gene knockout (KO) on C2C12 cell identity. a, b Influence of Myod1 loss on muscle differentiation in C2C12 cells, as viewed by the sharp contrast in morphological changes (a) and by IF detection of muscle-specific markers (b) in WT and KO cells after 7 days of culture in muscle differentiation medium that contained 2% horse serum (HS). c, d Evidence for the gain of properties of NSCs/NPCs in KO cells, as revealed by neurosphere formation by KO cells but not by WT cells after 6 days of culture in serum-free medium (c). IF detection exhibits also difference in expression of Mhc and Sox1 in these cells (d). In b and d, nuclei were counterstained with DAPI. e IB detection of proteins specific for embryonic neural cells in C2C12 WT and KO cells cultured in serum-free medium for 6 days. IB was done with whole cell lysates. β-Act was used as a loading control. f–h Transcriptomic profiling of WT and KO cells with RNA-sequencing. A RNA-seq heatmap (f) shows 3201 DEGs between WT and KO cells, both in triplicate. Red represents upregulated expression, while green means downregulated expression. g shows top 20 of enriched GO terms for DEGs, and h represents the numbers of DEGs according to KEGG gene classification