Fig. 6

IGF2-intron3-C3071T mutation improved adipogenesis through PI3K-AKT/AMPK pathway. A EdU and Ki67 immunofluorescent staining were performed between WT MSCs, ED MSCs, and ED MSCs with the treat of MK2206, AICAR, or MK2206 + AICAR. The percentage of EdU-positive or Ki67-positive cells were presented as histogram in the right panel. Scale bar = 100 μm. B Real-time cell proliferation monitoring assay was carried to measure the proliferation index of the above 5 groups, n = 3 per group. Mark of significance, *: WT vs. ED; a: ED vs. ED + MK2206; b: ED vs. ED + AICAR; c: ED + AICAR vs. ED + MK2206 + AICAR. Significance (P < 0.01) was reached from 30 to 60 h, only 60 h was marked in the figure. C Oil red O staining of lipid droplet formation at d 6 of induced adipogenic differentiation in the above 5 groups. The absorption value of triglyceride content was presented as histogram in the right panel. Scale bar = 100 μm. D qPCR analyzed the relative transcription levels of downstream target genes in AMPK signaling pathway. Mark of significance was same as B. E Schematic diagram of the regulatory mechanism of IGF2-intron3-C3071T mutation on improved myogenesis and adipogenesis. All data were presented as mean ± SEM. n = 3–4 per group. ^*P < 0.05, ^**P < 0.01, ^***P < 0.01, ns: P > 0.05, one-way ANOVA or two-way ANOVA