Fig. 6

Increase of injury-activated glial cell numbers in the wound bed boosts myofibroblast differentiation through TGF-β activation. a Scheme of the experimental strategy for Plp-CreER^T2-mediated glial cell expansion in our murine wound healing model through Pten cKO. b Immunolabelling of pAKT (green) and Sox10 (red) in NB of skin wound sections at D7 shows a significant increase in PI3K signalling upon Pten cKO. c Immunolabelling of p75 (green) in Ctrl and Pten cKO skin wound sections at D14. d, e Quantification of p75⁺ and tdTomato⁺ cells per mm² of granulation tissue shows a significant increase in p75⁺ cells at D7 and D14 and traced cells at D7 upon Pten cKO. Quantification of D7 wound sections of the percentage of f wound closure, g area of HPE, h wound contraction, i re-epithelialisation reveals no difference in these parameters. j Immunolabelling of α-SMA (green) in Ctrl and Pten cKO skin wound sections at D7. k, l Quantification of the percentage of α-SMA⁺ area of the granulation tissue at D7 and of the percentage of pSMAD2⁺ cells per α-SMA⁺ area of the granulation tissue of skin wounds at D5 shows increase of both myofibroblast-occupied area and canonical TGF-β signalling in Pten cKO skin wounds. Note that control data sets in Fig. 6k and Fig. 5i are the same because the experiments were performed at the same time. Data are represented as mean ± SEM of N = 2, n = 8 (Ctrl D7), N = 2, n = 8 (Ctrl D14); N = 3, n = 10 (Pten cKO D7), N = 2, n = 8 (Pten cKO D14) (d); N = 4, n = 7 (Ctrl), N = 2, n = 6 (Pten cKO) (e); N = 4, n = 16 (Ctrl), N = 5, n = 20 (Pten cKO) (f); N = 3, n = 12 (Ctrl), N = 4, n = 16 (Pten cKO) (g); N = 4, n = 16 (Ctrl), N = 6, n = 24 (Pten cKO) (h); N = 4, n = 16 (Ctrl), N = 5, n = 20 (Pten cKO) (i); N = 5, n = 19 (Ctrl), N = 5, n = 20 (Pten cKO) (k); N = 3, n = 8 (Ctrl), N = 2, n = 8 (Pten cKO) (l). Scale bars, 50 µm (b), 200 µm (c, j)