Fig. 6

Effects of IGF2 knockdown in XF-iMSCs on Col6a1-KO/NSG MuSC differentiation. a mRNA expression of IGF2 and PXDN. The mRNA expression level of each gene was analyzed using RT-qPCR in Ad-MSCs, BM-MSCs, and XF-iMSCs. Levels are shown relative to those in XF-iMSCs. Data are shown as the mean ± SD. b Concentration of IGF2 in the culture supernatants of Ad-MSCs, BM-MSCs, and XF-iMSCs as obtained using ELISA. Data are shown as the mean ± SD. c mRNA expression level and concentration of IGF2 in the culture on co-culture day 3. Data are presented as the mean ± SD. mRNA expression levels are shown with levels relative to those in XF-iMSCs. d Representative immunofluorescence images of Col6a1-KO/NSG mouse-derived MuSCs 3 days after single culture or co-culture with XF-iMSC, XF-iMSC_IGF2-KD, or XF-iMSC_mock. e Total number of DAPI + /hLamin A/C- mouse myogenic cells 3 days after co-culture. Data are expressed relative to XF-iMSCs and are presented as the mean ± SD of three independent experiments. f Percentage of Pax7 + /MyoD-, Pax7 + /MyoD + , and Pax7-/MyoD + cell populations 3 days after co-culture. Data from three independent experiments are shown as the mean ± SD. g mRNA expression level and concentration of IGF2 in the culture on co-culture day 6. Data are presented as the mean ± SD. mRNA expression levels are shown with levels relative to those in XF-iMSCs. h Representative immunofluorescence images of Col6a1-KO/NSG mouse-derived MuSCs 6 days after single culture or co-culture with XF-iMSCs, XF-iMSC_ IGF2-KD, or XF-iMSC_mock. i, j Area of MHC + myotubes (i) and number of MHC + myotubes (j) with four or more nuclei on day 6 after co-culture. Data are expressed relative to XF-iMSCs and are presented as the mean ± SD of three independent experiments. co-cluture experiment: n = 6 (XF-iMSCs, XF-iMSC_ IGF2-KD and XF-iMSC_mock), n = 3 (single culture). qPCR and ELISA experiment: n = 2 (Ad-MSCs, BM-MSCs, XF-iMSCs)