Fig. 2

MYH1G-AS is a skeletal muscle-specific lncRNA that is coordinately regulated by SMAD3 and SP2. A, B Relative MYH1G-AS expression in PEM and SOL of 7-week-old Xinghua chicken detected by RNA-seq (A) and qPCR (B). C Tissue expression profiles of MYH1G-AS. D Relative MYH1G-AS expression during CPM proliferation and differentiation. E The distribution of MYH1G-AS in the cytoplasm and nucleus of CPMs determined by qPCR. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and U6 serve as cytoplasmic and nuclear localization controls, respectively. F RNA in situ hybridization of MYH1G-AS in CPM. Special FISH probes against MYH1G-AS were modified by Cy3 (red). The nucleus was stained by DAPI (blue). G Western blot analysis of the coding ability of MYH1G-AS. CPMs transfected with β-actin were used as a positive control (PC) and untransfected CPMs were used as a negative control (NC). H Significantly enriched TFs in MYH1G-AS upstream the ATAC-seq peak was predicted by MEME suite. I Transcriptional activity of MYH1G-AS upstream ATAC-seq peak. J ChIP analysis of the binding capacity of SMAD3 to the MYH1G-AS promoter. K, L Relative SMAD3 (K) and MYH1G-AS (L) expression after SMAD3 overexpression or interference. M ChIP analysis of the binding capacity of SP2 to MYH1G-AS promoter. N, O Relative SP2 (N) and MYH1G-AS (O) expression with SP2 overexpression or inhibition. Results are presented as mean ± SEM. In panels A–D and I–O, statistical significance of differences between means was assessed using an independent sample t-test. (*P < 0.05; **P < 0.01)