Fig. 7

The association between Msx1 and Emerin is necessary for inhibition of myoblasts’ differentiation and the repression of myogenic targets. a Western blotting assays. To ensure Flag-Msx1 was successfully overexpressed and Emerin was efficiently knockdown in C2C12 myoblasts used in real-time PCR (b) and IF assays (c). (b) mRNA levels of Msx1 target myogenic genes MyoD and Myf5. Data were shown as the relative expression levels in cells overexpressing Msx1 or the control vectors. Values were the mean ± SD. ***p < 0.0001, **p < 0.001, *p < 0.01. c IF assays. C2C12 myoblasts were transfected with epiCRISPR/Cas9 plasmids firstly to acquire Emerin deficient cells (transfected with Cas9-gEmerin) and control cells (transfected with Cas9). Then these cells were infected by retrovirus with plasmids expressing Flag-Msx1 and GFP (vectors overexpressing Msx1) or only GFP (control vectors). IF assays were performed after induction by DMEM with 2% horse serum for 3–5 days as detected using antibody for myosin heavy chain (MHC) and DAPI. Cells expressing GFP were successfully infected by retrovirus. Scale bars represented 200 µm. d Schematic diagram of Ship-to-Anchor model. Emerin associated with Msx1 at the nuclear periphery to help Msx1 with the recruitment of histone methyltransferase Ezh2 to target genes