Fig. 2: RNA-seq and proteomics analysis of foetal skeletal muscle and verification.

A TEM showed myofiber and mitochondrial malformation in foetal GDM lower limb skeletal muscle; B Principal component analysis of proteomics verified that protein sample between groups were sufficiently heterogeneous; C DEPs were mainly distributed in nucleus, cytosol and mitochondria; D KEGG analysis of DEPs showed OXPHOS and TCA cycle were highly enriched; E Volcano plot of DEGs showed 993 upregulated and 653 downregulated genes were identified as significance; F Histogram of gene transcription levels related to mitochondria and metabolic processes according to RNA-seq; G Venn diagram of coanalysis of DEGs in RNA-seq and DEPs in proteomics; H, I GO analysis showed both upregulated DEPs and DEGs were mainly enriched in positive gene expression and transcription; J GO analysis showed both downregulated DEPs and DEGs were mainly enriched in mitochondrion and fatty-acid or lipid metabolic process; K KEGG analysis of both downregulated DEPs and DEGs were mainly focused on PPAR signalling and thermogenesis; L IPA software predicted that Ppargc1α inhibition was the key regulator of mitochondrial and oxidative disorder; M RT-qPCR showed representative mRNA levels related to mitochondrial biogenesis and fatty acid oxidation in foetal muscles were largely inhibited in GDM male offspring (n = 5, multiple t tests); N RT-qPCR showed mRNA levels of Ppargc1α decreased at ED18.5 (n = 5), 6 weeks (n = 3) and 8 weeks (n = 4) in GDM male offspring by multiple t tests; Data are expressed as the mean ± SEM. Significance of the differences: *p < 0.05, **p < 0.01, ***p < 0.001.