Journal: Bone Research
Article Title: TGF-β-induced fibrotic scar formation limits recovery of spinal cord injury
doi: 10.1038/s41413-026-00507-7
Figure Lengend Snippet: Conditional knockout of Tgfβ1 in macrophage lineage cells reduces pSmad2 levels and promotes functional recovery in SCI mice. a – d ELISA analysis showing the concentration of TGF-β in the spinal cord and the serum after SCI in each mouse for the groups indicated (* P < 0.05, ** P < 0.01, n = 4). e Representative Western blots showing the activation of phosphorylated Smad (pSmad) signaling. f Representative images of immunofluorescent staining of pSmad2 + (red), platelet-derived growth factor receptor-β + (PDGFR-β + ) (green) pericytes , and DAPI (blue) staining of nuclei at 7 days after SCI. Scale bar, 10 µm. g Representative immunofluorescence images of PDGFR-β + (green) pericytes and DAPI (blue) staining of nuclei at 2 weeks after SCI. Scale bars, 200 µm. h Quantitative analysis of the number of pSmad2 + cells (**** P < 0.000 1, n = 6). i Quantitative analysis of the number of PDGFR-β + cells (** P < 0.01, n = 6). j , k Illustrations of the hot plate test and the von Frey test. l Quantitative analysis of hindpaw withdrawal time responding to temperature (hot plate test, ** P < 0.01, n = 8). m , n Quantitative analysis of hindpaw withdrawal frequency responding to mechanical stimulation (von Frey test, 0.7 mN and 3.9 mN, * P < 0.05, ** P < 0.01, n = 8). o Quantitative analysis of Basso Mouse Scale (BMS) score between Tgfb1 flox / flox control mice after SCI, Tgfb1 LysM-cre −/− mice after SCI, and Tgfb1 flox/flox control mice without SCI (* P < 0.05, n = 8). PWF paw withdrawal frequency. LF left forepaw, RF right forepaw, RH right hindpaw. Statistical significance was determined by multifactorial ANOVA, and all data are shown as means ± standard deviations. Source data are provided as a Source Data file
Article Snippet: The sections were incubated with primary antibodies to rabbit 5-HT (1:50, sc-65495, Santa Cruz Biotechnology, Dallas, TX, USA), mouse β-III-tubulin (1:100, MA1-118, Invitrogen, Carlsbad, CA, USA), rabbit PGP9.5 (1:250, ab108986, Abcam, Cambridge, UK), mouse PGP9.5 (1:50, ab8189, Abcam), rabbit Fibronectin (1:100, ab2413, Abcam), mouse Fibronectin (1:100, ab6328, Abcam), rabbit Collagen III (1:100, ab7778, Abcam), chicken GFAP (1:500, ab4674, Abcam), mouse Collagen1α1 (1:50, sc-293182, Santa Cruz Biotechnology), rabbit Phospho SAMD2 (1:100, 44-244 G, Invitrogen), rabbit PDGFR-β (1:100, ab32570, Abcam), goat PDGFR-β (1:100, AF1042, R&D Systems), rabbit FSP1 (1:300, 07-2274, MilliporeSigma, Burlington, MA, USA), and chicken green fluorescent protein (1:250, ab13970, Abcam) overnight at 4 °C.
Techniques: Knock-Out, Functional Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Western Blot, Activation Assay, Staining, Derivative Assay, Immunofluorescence, Hot Plate Test, Control