Fig. 3

There is no difference in the transcriptional activity of MyoD among four species. a Alignment of bHLH domain among mouse, pig, chicken and xenopus. b qPCR analysis for the expression of myoD in NIH3T3 cells treated with overexpression Flag-myoD plasmid of different species or a control Flag vector in GM for 36 h. c After transfection of 48 h, MyoD protein level was detected by Flag antibody. Ctrl represents a control group. (d, e) qPCR analysis was performed to detect the expression level of myog, ckm, cdh15 and myh3 in different species at 3d (d) or 6d (e) post-differentiation. f, g The significance analysis of expression levels of key factors of myogenic differentiation from D and E in different species to determine whether MyoD has the same transcriptional activity. h 293 T cells were transiently transfected with 4Rtk-luc and myoD expression vectors of different species. After transfection of 48 h, cells were harvested to measure Firefly and Renilla luciferase activities by Dual-Luciferase Reporter Assay System Kit (Promega). i Immunofluorescence detection of MyHC (green) in myoD plasmid of mouse, pig, chicken and xenopus or control transfected NIH3T3 cells after 7 days of differentiation. Data are presented as mean ± S.E.M.; n = 3; *p < 0.05, **p < 0.01, ***p < 0.001 (Student’s t-test); Scale bar =100 μm