Figure 1: Incomplete regeneration following deletion of Numb:Numbl in satellite cells.
From: Numb is required to prevent p53-dependent senescence following skeletal muscle injury
(a) Strategy used to study TA muscle regeneration in the context of Numb:Numbl depleted satellite cells. Data were collected from transverse TA muscle cryosections of adult mice. (b) Histological staining with H&E. (c) Histological staining with Von Kossa. Dark precipitates reveal calcium deposits, a sign of chronic inflammation. (d) Immunofluorescence staining for macrophages (arrows) using anti-F4/80 antibody. (e) Quantification of the number of macrophages in (d); control: n=5 mice, 3.2±0.4 cells per unit area; mutant: n=5 mice, 5.6±0.4 cells per unit area; Mann-Whitney (MW) test P=0.079. (f) Histological staining with Sirius Red. (g) Quantification of fibrosis based on Sirius Red histology in (f); control: n=5 TA, 5 mice, 5.6±0.3% fibrosis per unit area; mutant: n=5 TA, 5 mice, 12±3.5% fibrosis per unit area; MW test P=0.0397. (h) Immunofluorescence staining for fibroblasts (arrows) using anti-TCF4 antibody. (i) Quantification of the number of fibroblasts in (h); control: n=4 mice, 14.2±1.4 cells per unit area; mutant: n=4 mice, 18.8±1.12 cells per unit area; MW test P=0.0571. (j) Quantification of the numbers of mGFP+ fibres; control: n=3 TA, 3 mice mice, 45±3.1 fibres per unit area; mutant: n=6 TA, 3 mice, 41.4±3.7 fibres per unit area; MW test P=0.2619. (k) Immunofluorescence staining for quiescent satellite cells (arrows) using anti-Calcitonin receptor antibody. (l) Quantification of quiescent satellite cells in (k); control: n=4 mice, 2.17±0.15 cells per unit area; mutant: n=4 mice, 1.83±0.14 cells per unit area; MW test P=0.20. (m) Quantification of the number Pax7nGFP+ cells in one diaphragm for quiescent cells; control: n=8 mice, 60,376±4,038; mutant: n=7 mice, 65,663±10,106; MW P=0.6943) and in 2 TA for activated cells (control 5 DPI: n=3 mice, 174,880±15,190; mutant 5 DPI: n=3 mice, 170,490±4,747, MW P>0.99; control 8 DPI: n=12 mice, 74,306±5,002; mutant 8 DPI: n=7 mice, 77,852±9,562, MW P=0.9671; control 21 DPI: n=3 mice, 59,935±5,284; mutant 21 DPI: n=4, 36,867±4,276, MW P=0.0571). Quantifications are presented as mean±s.e.m. Scale bars b,c,f: 100 μM; d,h: 25 μM; k: 20 μM. NS, not significant.
