Extended Data Fig. 3: COLV delays proliferation and differentiation of satellite cells.
From: Reciprocal signalling by Notch–Collagen V–CALCR retains muscle stem cells in their niche
a, Experimental scheme: isolated Tg:Pax7-nGFP satellite cells cultured overnight (o/n) before collagen treatment. b, Myosin heavy chain (MyHC) and EdU staining of satellite cells treated with COLI or COLV. Fusion index: 82%, 86% and 33% for HOAc solvent, COLI and COLV, respectively (n = 3 mice, ≥250 cells, 2 wells per condition). c, Percentage of EdU+ primary myogenic cells after ten days of culture with indicated collagens. EdU: 2.6%, 1.3% and 18.2% for COLI, COLVI and COLV, respectively (n = 3 mice, ≥250 cells, 2 wells per condition). d, Experimental scheme for control and cKO mice. Satellite cells were plated overnight before collagen treatment. e, GFP and MyHC immunostaining of Rbpj cKO satellite cells (n = 3 mice per condition) incubated 60 h in presence of COLI or COLV, or with HOAc control (n = 3 mice, ≥200 cells, 2 wells per condition). f, Percentage of EdU+ cells (2 h pulse) of Rbpj-null primary myogenic cells, after ten days of culture with HOAc or indicated collagens. EdU: 1.0% and 7.6% for COLI and COLV, respectively (n = 3 mice, ≥150 cells, 2 wells per condition). g, RT–qPCR on GFP+Rbpj-null satellite cells isolated by FACS and cultured for 72 h in the presence of COLI or COLV. Results are normalized to Tbp. Data are mean ± s.d.; two-sided paired t-test; P value: two-sided unpaired t-test. Scale bars, 50 μm.
