Fig. 2: LncMDP1 promotes the proliferation and differentiation of myoblasts.

a RNA-FISH of lncMDP1. The scale is 10 μm. b LncMDP1 is located in the cytoplasm and nucleus, n = 3 biologically independent samples. c RNA expression levels of lncMDP1 in leg muscle at different times. Among them, E10, E12, E14, E16, E18 and 1d are n = 3 biologically independent samples, and 1w, 3w and 5w are n = 4 biologically independent samples. d Relative expression of CDK1, PCNA, CCND1, CCNB2, and P21 48 h after the transfection of si-lncMDP1 in proliferating myoblasts, n = 3 biologically independent samples. e LncMDP1 significantly reduced the protein expression level of CDK1 after interference, n = 3 biologically independent samples. The error bars are equivalent throughout the Figure. In the box line, a line in the middle of the box represents the median, and the upper and lower bottoms of the box are the upper quartile and the lower quartile, respectively. The blue circle represents si-NC, and the orange circle represents si-lncMDP1. Western blot was performed on different membranes for proteins with a molecular weight difference of fewer than 5 kDa (similar protein size, difficult to distinguish), and the sample size was consistent under the premise of detecting protein concentration. In this case, the sample and the loading control ran on different gels and thus transferred to different membranes. f Cell cycle analysis after the transfection of si-lncMDP1 in myoblasts, n = 3 biologically independent samples. g CCK-8 was used to detect the effect of lncMDP1 interference on myoblast proliferation, n = 5 biologically independent samples, among them, si-NC at 12 h and 24 h n = 6. h EdU staining was performed to analyze the effect on the proliferation of myoblasts after transfection with si-lncMDP1 and si-NC at a rate of 100 μm (si-NC is n = 4 biologically independent samples, and si-lncMDP1 is n = 3). i LncMDP1 interference significantly reduced RNA expression levels of myoblast differentiation-associated marker genes, including MyoD (n = 4 biologically independent samples), MyoG (si-NC is n = 3 biologically independent samples, and si-lncMDP1 is n = 4), MyHC (n = 3 biologically independent samples), and Myomarker (n = 3 biologically independent samples). j Knockdown of lncMDP1 reduced the protein expression levels of MyHC, n = 3 biologically independent samples. The error bars are equivalent throughout the Figure. In the box line, a line in the middle of the box represents the median, and the upper and lower bottoms of the box are the upper quartile and the lower quartile, respectively. The blue circle represents si-NC, and the orange circle represents si-lncMDP1. k Immunofluorescence of myoblasts differentiated with si-lncMDP1 and si-NC for 48 h, followed by staining with an MyHC antibody, n = 3 biologically independent samples. The scale is 100 μm. The results are shown as the mean ± SEM of three independent experiments. (*P < 0.05; **P < 0.01, ***P < 0.001).