Extended Data Fig. 4: COLV—and specifically α3-COLV—is critical for satellite cell self-renewal.
From: Reciprocal signalling by Notch–Collagen V–CALCR retains muscle stem cells in their niche
a, RT–qPCR of Col5a1 in control (Ctr; Tg:Pax7-CT2;Col5a1+/+;R26mTmG), heterozygous (Het; Tg:Pax7-CT2;Col5a1flox/+;R26mTmG) and conditional knockout (cKO; Tg:Pax7CT2;Col5a1flox/flox;R26mTmG) mice two weeks after tamoxifen diet (n = 3 mice per genotype). b, Transcript levels of the different Col5 mRNA chains in C2C12 after transfection of either control scramble, Col5a1 or Col5a3 siRNA, showing the specificity of each siRNA for its given targeted mRNA. Data are normalized to Tbp gene expression (n = 3 independent assays). c, Col5a1 and Col5a3 siRNA transfection of Tg:Pax7-nGFP isolated single myofibres cultured for 72 h and immunostained for GFP and MYOD. Resident satellite cells enter the myogenic program and form clusters composed of proliferating (PAX7+MYOD+MYOG−), differentiated (PAX7−MYOG+) and self-renewed (PAX7+MYOD−) cells within 72 h. Quantification of PAX7+MYOD−, PAX7+MYOD+ and PAX7−MYOD+ populations 72 h after transfection. Scramble siRNA was used as negative control (n ≥ 15 fibres counted from 3 mice). Data are mean ± s.d.; a, two-sided unpaired t-test; b, c, two-sided paired t-test. Scale bar, 50 μm.
