Fig. 4
From: 3D-environment and muscle contraction regulate the heterogeneity of myonuclei
BMP signalling regulates the incorporation of fibroblast nuclei into myotubes. A Fluorescent in situ hybridization to longitudinal sections of limb muscles with BMP4 probe (red) followed by an immunohistochimistry with the MF20 antibody (myosins, green) combined with DAPI staining (nuclei, blue). B,B’ Immunohistochemistry to adjacent longitudinal sections of limb muscle sections with pSMAD1/5/9 (red) and MF20 (myosins, green) antibodies combined with DAPI staining (nuclei, blue). C Immunohistochemistry to transverse sections of limb muscles with pSMAD1/5/9 (red) and MF20 (myosins, green) antibodies). D,D’,D” Immunohistochemistry to quail myoblast cultures with pSMAD1/5/9 (red), PAX7 (grey) and MF20 (myosins, green) antibodies combined with DAPI staining (nuclei, blue). E-J BMPR1Aca-transfected chicken fibroblasts (E), BMP4-transfected chicken fibroblasts (F), BMPR1Adn-transfected chicken fibroblasts (H) or NOGGIN-transfected chicken fibroblasts (I) were co-cultured with quail myoblasts; and labelled with the QCPN (quail nuclei, red), MF20 antibody (myosins, green) combined with DAPI staining (nuclei, blue). G Percentage of chicken fibroblast myonuclei within myotubes in control-, BMPR1Aca-, BMP4-transfected chicken fibroblasts cultured with quail myoblasts, (BMP gain-of-function experiments). J Percentage of chicken fibroblast myonuclei within myotubes in control-, BMPR1Adn-, NOGGIN-transfected chicken fibroblasts cultured with quail myoblasts, (BMP loss-of-function experiments). (G,J) Graphs shows mean ± s.d