Fig. 5
IGF2-intron3-C3071T mutation promoted cell proliferation, myogenesis and adipogenesis of MSCs isolated from the LL muscle of 7-day-old female piglets. A and B The mRNA expression and protein levels of IGF2 were detected in MSCs. n = 3 per group. C The top 20 of KEGG pathways enriched by transcriptomic analysis in MSCs. D Heatmaps of IGF2, and key genes involved in myogenesis and adipogenesis. E and F EdU and Ki67 immunofluorescent staining were performed between IGF2C/T and WT MSCs, the percentage of EdU-positive or Ki67-positive cells were presented as histogram in the right panel. Scale bar = 100 μm. n = 3 per group. G Real-time cell proliferation monitoring assay of the proliferation index between IGF2C/T and WT MSCs. n = 8 per group. H qPCR analyzed the mRNA levels of related genes in the proliferation phase. n = 3 per group. I Immunofluorescent staining of MyHC was performed to detect myotube formation at d 4 of induced myogenic differentiation. The fusion index was presented as histogram in the right panel. Scale bar = 100 μm. n = 3 per group. J qPCR analyzed the mRNA levels of related genes at d 4 of induced myogenic differentiation. n = 3 per group. K Oil red O staining of lipid droplet formation at d 6 of induced adipogenic differentiation. The absorption value of triglyceride content was presented as histogram in the right panel. Scale bar = 50 μm. n = 3 per group. L Immunofluorescence co-staining of perilipin and bodipy between IGF2C/T and WT MSCs at d 6 of induced adipogenic differentiation. Scale bar = 50 μm. n = 3 per group. M qPCR analyzed the mRNA levels of related genes at d 6 of adipogenic differentiation. n = 3 per group. N Western blot analyzed the protein levels of AKT, AKT-S473, AMPKα, AMPKα-T172 and SREBP1 at d 6 of induced adipogenic differentiation. All data were presented as mean ± SEM. *P < 0.05, **P < 0. 01, ***P < 0.001, student's t-test