Fig. 5: Discrete beneficial functions of early myofibroblasts and late lymphocyte-interactive fibroblasts.

a,b, tMCAO-induced fibrotic lesion at 14 dpi (a) and mouse survival (b). Control stroke: n = 18, Tgfbr2-cKO stroke: n = 21, control sham/rest: n = 10, Tgfbr2-cKO sham/rest: n = 15 mice. Mice were treated with tamoxifen at −3 to −1, 2, 5, 8 and 11 dpi. Scale bars: 500 μm (main images), 200 μm (enlarged views). c–f, Ipsilateral hemisphere area (c), brain midline shift (d), oligodendrocyte density (e) and FluoroJade-C⁺ (degenerating) neuron density (f) at 3 days after tMCAO. Control, n = 4, Tgfbr2-cKO, n = 6 mice. Two slices per mouse (c,d); light grey or red dots and text represent values and P values per tissue slice, dark colors per mouse (c); normalized to contralateral (c,e,f, per group). g,h, Heart rate (g) and mean arterial pressure (h) 1 to 3 days after tMCAO. Control groups: n = 7, Tgfbr2-cKO groups: n = 4 mice (g); control uninjured: n = 8, Tgfbr2-cKO uninjured: n = 5, control stroke: n = 7, Tgfbr2-cKO stroke: n = 4 mice (h). n = 1 Tgfbr2-cKO stroke recorded at 1 dpi, n = 1 Tgfbr2-cKO stroke recorded at 2 dpi; remaining mice recorded at 3 dpi. i, Schematic showing loss of CXCL12 production in Col1a2^creER; Cxcl12^flox fibroblasts with tamoxifen treatment at 0 to 2,7 to 9 and 14–16 dpi. j–l, Surfaced T cells (CD3ε⁺CD45⁺, processed via Imaris software ‘surface’ function) near lymphocyte-interactive fibroblasts (CD80⁺) in control (j) or Cxcl12-cKO mice at 21 dpi (k) with quantification (l). n = 7 mice per group; 2 slices per mouse. Scale bars: 500 μm (left), 100 μm (right). m–p, IFNγ expression in CD8⁺ T cells (m), IFNγ (n) and IL-17A (o) expression in CD4⁺ conventional T (T_conv) cells, and IL-17A expression in γδ T cells (p) at 21 dpi. Control: n = 11, Cxcl12-cKO: n = 14 mice. q, Cortical neutrophils in control and Cxcl12-cKO mice at 21 dpi. Control, n = 21, Cxcl12-cKO: n = 16 mice. r, Schematic of snRNA-seq experiment. Control (wild-type (WT)) and Cxcl12-cKO mice were collected at 21 dpi (2 mice per genotype), lesions and parenchyma were micro-dissected and multiplexed, and nuclei were sorted and sequenced (19,668 nuclei). s, Global UMAP analysis of samples depicted in r. t, Type 1 T cell abundance (180 nuclei). u, IFNγ response score among lesional myeloid cells. Control: n = 1,659, Cxcl12-cKO: n = 2,056 nuclei. v, UMAP of neuronal cells (5,862 nuclei). w, IFNγ response score of excitatory and inhibitory neurons. Wild-type excitatory: n = 2,392, wild-type inhibitory: n = 576, Cxcl12-cKO excitatory: n = 2,229, Cxcl12-cKO inhibitory: n = 665 nuclei. x, Schematic showing a model of T cell regulation via late lesional fibroblast-derived CXCL12. log-rank (Mantel–Cox) test, Bonferroni correction (k = 4) (b); two-way Student’s t-test (c–e,l–p); two-way repeated-measures ANOVA, Sidak’s post-test (f,q (repeated-measures),g,h); two-way Mann–Whitney test (u,w (Bonferroni correction)). Slice thickness: 14 μm. Images represent two or more mice.

Source data