goat anti pdgfr β  (R&D Systems)

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

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 TGF-β type 2 receptor ( Tgfbr2 ) in pericytes reduces fibrotic scar formation in SCI mice. a Schematic diagram of Tgfbr2 knockout in Glast-Cre + pericytes. b Representative images of immunofluorescent analysis of PGP9.5 + (red) nerve fibers, fibronectin + (green) fibrotic scar, and DAPI (blue) staining of nuclei at 4 weeks after SCI. Scale bar, 200 µm. c , d Quantitative analysis of the intensity value of PGP9.5 and fibronectin (* P < 0.05, n = 6). e Representative images of immunofluorescent analysis of nerve axon–specific β-III-tubulin + (green), collagen III + (red) fibrotic scar, and DAPI (blue) staining of nuclei at 4 weeks after SCI. Scale bar, 200 µm. Right images are high resolution versions of the boxed regions in the left images. Scale bar, 50 µm. f Quantitative analysis of the intensity value of collagen III (* P < 0.05, n = 6). g Representative images of immunofluorescent analysis of neurotransmitter marker 5-HT + (red), nerve axon–specific β-III-tubulin + (green), and DAPI (blue) staining of nuclei in spinal cord lesion site of T10 in Tgfbr2 flox/flox sham group mice, Tgfbr2 flox/flox control mice, and Tgfbr2 Glast-creER −/− mice at 4 weeks after SCI. Scale bar, 200 µm. Right images are high-resolution versions of the boxed regions in the left images. Scale bar, 50 µm. h , i Quantitative analysis of the intensity value of 5-HT and β-III-tubulin (* P < 0.05, n = 6). j Representative images of immunofluorescent analysis of pSmad2 + (red), PDGFR-β + (green) pericytes and DAPI (blue) staining of nuclei at 7 days after SCI. Scale bar, 10 µm. k , l Quantitative analysis of the intensity mean value of PDGFR-β and the number of pSmad2 + cells (*** P < 0.001, **** P < 0.000 1, n = 6). m Representative Western blots showing the activation of pSmad signaling. 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, Staining, Marker, Control, Western Blot, Activation Assay

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: LepR + MSCs are the primary cells for fibrotic scar formation. a , b Diagram illustrating the genetic strategy to trace type A pericytes and the timeline of tamoxifen injection. c Representative images of immunofluorescent analysis of tdT + (red) type A pericytes, PDGFR-β + (green) pericytes, and DAPI (blue) staining of nuclei in the spinal cord lesion site of T10 wild-type (WT) mice at 2 weeks after SCI. Scale bar, 50 µm. Left images are high-resolution versions of the boxed regions in the right images. Scale bar, 10 µm. d Quantitative analysis of percentage of PDGFR-β cells/tdTomato + type A pericytes cells (*** P < 0.001, n = 4). e Representative images of immunofluorescent analysis of collagen III + (Col-III green) fibrotic scar, tdT + (red) type A pericytes, and DAPI (blue) staining of nuclei in spinal cord lesion site of T10 in 13C4 group control mice and 1D11 group mice at 4 weeks after SCI. Scale bar, 50 µm. f Quantitative analysis of percentage of collagen III + area /tdTomato + type A pericytes field (*** P < 0.001, n = 4). g Representative images of immunofluorescent analysis of LepR + (green) MSCs, tdT + (red) type A pericytes, and DAPI (blue) staining of nuclei in sham group mice, 13C4 group control mice, and 1D11 group mice at 4 weeks after SCI. Scale bar, 50 µm. h Quantitative analysis of the percentage of LepR + cells/tdTomato + cells (* P < 0.05, n = 4). i Representative images of immunofluorescent analysis of LepR + (green) MSCs, collagen III + (Col-III, green) fibrotic scar , and DAPI (blue) staining of nuclei. Scale bar, 50 µm. j Quantitative analysis of the percentage of collagen III + area/ LepR + MSCs field (** P < 0.01, n = 4). k Representative images of immunofluorescent analysis of LepR + (green) MSCs, FSP1 + (red) type A pericytes , and DAPI (blue) staining of nuclei. Scale bar, 50 µm. l Quantitative analysis of the percentage of LepR + MSCs/FSP1 + cells (** P < 0.01, n = 4). 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: Injection, Staining, Control

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: Neonatal mice completely recover from SCI without scarring. a Schematic diagram illustrating the timeline of the experimental procedures. b Representative images of immunofluorescent analysis of pSmad2 (red), PDGFR-β (green), and DAPI (blue) in the spinal cord lesion site in neonatal mice on days 2, 7, and 12 with or without SCI. Scale bar, 50 µm. Right images are high-resolution versions of the boxed regions in the left images. Scale bar, 50 µm. c , d Quantitative analysis of the number of pSmad2 + cells and PDGFR-β + cells (** P < 0.01, *** P < 0.001, **** P < 0.000 1, n = 6). e Representative images of immunofluorescent analysis of β-III-tubulin (green), collagen III (red), and DAPI (blue) in the spinal cord lesion site in neonatal mice on days 2 and 12, and adult mice with or without SCI. Scale bar, 200 µm. f , g Quantitative analysis of the intensity mean value of collagen III + fibrotic scar and β-III-tubulin + nerves (* P < 0.05, **** P < 0.000 1, n = 6). 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: