Fig. 4: Brain lesional myofibroblasts are coordinated by TGFβ signalling, profibrotic myeloid cells and perilesional glia to drive wound healing and limit chronic inflammation.
From: Dynamic fibroblast–immune interactions shape recovery after brain injury
a–c, Treatments with control or clodronate (Clod) liposomes and images of fibroblasts and astrocytes in lesions (a), ECM (ER-TR7) coverage (b) and lesion size (c) at 14 dpi. b, Control: n = 7, clodronate: n = 6 mice. c, Control: n = 15, clodronate: n = 10 mice. Two slices per mouse. Scale bars, 500 μm. d, Schematic of ablated fibroblast TGFβ signalling (Col1a2creER; Tgfbr2flox; tamoxifen treatment on days −14 to −12, −7, 0 to 2, 5, 8 and every 3 days subsequently until sample collection). e–h, Images of control and Tgfbr2-cKO (cKO) lesions with thresholded ER-TR7 (e) or Col1a2creER; Rosa26tdT+ fibroblasts (f), ER-TR7 coverage (g) and lesion size (h) at 14 dpi. Control (ctrl): n = 20, Tgfbr2-cKO: n = 12 mice; 2 slices per mouse. Scale bars, 500 μm. i, Cortical neutrophils in controls and Tgfbr2-cKO lesions. 0 dpi control (non-littermate-sham): n = 8, 2, 4 and 7 dpi control and 2, 7 and 21 dpi Tgfbr2-cKO: n = 6, 14 dpi control: n = 12, 21 dpi control: n = 9, 0 dpi Tgfbr2-cKO: n = 3, 4 dpi Tgfbr2-cKO: n = 5, 14 dpi Tgfbr2-cKO: n = 7 mice. j–l, Images of lesions in control (cre-negative or vehicle) and myofibroblast deleter (Cthrc1creER; Rosa26DTA) mice (j), and ER-TR7 coverage (k) and lesion size (l) at 21 dpi. Control: n = 19, deleter: n = 22 mice; 2 slices per mouse. Scale bars, 200 μm. m, Lesion size at 28 dpi in controls and Tgfbr2-cKO mice after late tamoxifen treatment (days 14–16, 19, 22 and 25). Control: n = 6, Tgfbr2-cKO: n = 7 mice per group; 2 slices per mouse. n, Cortical neutrophils at 21 dpi after late or continuous tamoxifen treatment. Control late: n = 7, Tgfbr2-cKO late: n = 6, Tgfbr2-cKO continuous: n = 6, control continuous: n = 9 mice. o–q, Images of cortical lesions in control (IgG) mice or mice with αvβ8 blockade (ADWA11) at 14 dpi (o), ER-TR7 coverage (p) and cortical neutrophils (q). IgG: n = 11, ADWA11: n = 9 mice (p; 2 slices per mouse); IgG: n = 7, ADWA11: n = 8 mice (q). Scale bars, 500 μm. r,s, Cortical lesions in control and GFAPcre; Itgb8flox mice at 14 dpi (r) and ER-TR7 coverage (s). Control: n = 12(control), Tgfbr2-cKO: n = 7 mice; 2 slices per mouse. Scale bars, 200 μm. t, Fibroblast snRNA-seq cluster abundance in control, Tgfbr2-cKO (Col1a2creER; Tgfbr2flox), and ADWA11-treated mice at 7 dpi (early) and 21 dpi (late). u, Expression of myofibroblast genes in myofibroblasts across time points. v–x, Myeloid cluster abundance (v), SAM frequency (flow cytometry) (w; 14 dpi) and perilesional T cell numbers (x; 14 dpi). n = 4 mice per group (w); control: n = 9, Tgfbr2-cKO: n = 6 mice (x; 2 slices per mouse). Quantification normalized to controls in each experiment (c,h,k,l). Two-way Student’s t-test (b,c,g–i (0 dpi), k–m,p,s,w,x); two-way repeated-measures ANOVA, Sidak’s post-test (i) (7–21 dpi, per time point; bold: lesioned control versus lesioned Tgfbr2-cKO, grey: lesioned control versus contralateral control, italic: lesioned Tgfbr2-cKO versus contralateral Tgfbr2-cKO); two-way repeated-measures ANOVA, Sidak’s post-test (q); two-way ANOVA, Sidak’s post-test (n). Slice thickness: 14 μm. Dotted lines indicate lesion boundary (a,e,f,j,o,r); images represent two or more mice.
