tunel staining promega g7130  (Promega)

Journal: Advanced Science

Article Title: Inhibition of Integrin α v β 6 Activation of TGF‐ β Attenuates Tendinopathy

doi: 10.1002/advs.202104469

Figure Lengend Snippet: CED mice show an Achilles tendon tendinopathy phenotype. A) Concentration of active TGF‐ β 1 in Achilles tendons of CED and WT mice. B) Message RNA expression of Scx in Achilles tendons of CED and WT mice. C) TUNEL staining and D) quantitative analysis of apoptotic cells in tendons. Scale bar: 50 µm. E) SOFG staining of normal tendon and CED mouse tendons. Proteoglycan (red) and collagenous matrix (green). Scale bar: 200 µm. F) Immunohistochemical staining and G) quantitative analysis of Col2 + chondrocytes per tissue area (mm 2 ) in Achilles tendons. Scale bar: 100 µm. H) Immunofluorescent staining of CD31 + cells and I) quantification of the fold change of blood vessels in CED mice normalized to that of WT mice in the Achilles tendons. Scale bar: 100 µm. J) Quantitative analysis of maximum tensile force and stiffness of Achilles tendons from WT and CED mice. K) Immunostaining of Col2 + cells (red) and yellow fluorescent protein (YFP) – positive cells (green) in Scx‐creERT2::EYFP mice 8 weeks after DI. Scale bar: 50 µm. L) Ratio of YFP + and YFP − cells in Col2 + cells in tendinopathic tendon 8 weeks after DI in Scx‐creERT2::EYFP mice. All data are shown as the mean ± standard deviation ( n = 8 per group). * p < 0.05 compared with WT littermates as determined by unpaired, 2‐tailed Student's t ‐test.

Article Snippet: Sections were processed for TUNEL staining according to the manufacturer's protocol (Promega, Madison, WI, catalog no. G3250, for Figures , , , and ; Promega, catalog no. G7360, for Figure ).

Techniques: Concentration Assay, RNA Expression, TUNEL Assay, Staining, Immunohistochemical staining, Immunostaining, Standard Deviation

Journal: Advanced Science

Article Title: Inhibition of Integrin α v β 6 Activation of TGF‐ β Attenuates Tendinopathy

doi: 10.1002/advs.202104469

Figure Lengend Snippet: Inhibition of TGF‐ β signaling attenuates tendinopathy. A–K) TGF‐ β neutralizing antibody attenuates tendinopathy. Mice are treated with 5 mg kg −1 body weight of the TGF‐ β neutralizing antibody 1D11 or control antibody 13C4 once per week for 8 weeks. A) Immunohistochemical staining and B) quantitative analysis of pSmad2 + cells per tissue area (mm 2 ) in Achilles tendons. Scale bar: 50 µm. C) SOFG staining of tendons in 13C4‐ or 1D11‐treated mice. Proteoglycan (red) and collagenous matrix (green). Scale bar: 50 µm. D) Frequency distribution of Achilles tendon orientation degree in 13C4‐ or 1D11‐treated mice. E) Immunohistochemical staining and F) quantitative analysis of Col2 + chondrocytes per tissue area (mm 2 ) in Achilles tendons. Scale bar: 100 µm. G) TUNEL staining and H) quantitative analysis of apoptotic cells in tendons. Scale bar: 100 µm. I) Immunofluorescent staining of CD31 + cells and J) quantification of the fold change of blood vessels in 1D11‐treated mice normalized to that of 13C4‐treated mice in the Achilles tendons. Scale bar: 100 µm. K) Quantitative analysis of maximum tensile force and stiffness of Achilles tendons measured 8 weeks after DI operation with 13C4 or 1D11 treatment. All data are shown as the mean ± standard deviation ( n = 8 mice per group). * p < 0.05 compared with 13C4 treated mice as determined by unpaired, 2‐tailed Student's t ‐test. L–V) Inducible knockout of Tgfbr2 in Scx + cells attenuates tendinopathy in mice. L) Immunohistochemical staining and M) quantitative analysis of pSmad2 + cells per tissue area (mm 2 ) in Achilles tendons. Scale bar: 50 µm. N) SOFG staining of tendons in Scx‐creERT2::Tgfbr2 f/f mice or its Tgfbr2 f/f littermates 8 weeks after DI. Proteoglycan (red) and collagenous matrix (green). Scale bar: 50 µm. O) Frequency distribution of Achilles tendon orientation degree in Scx‐creERT2::Tgfbr2 f/f mice or its wildtype controls. P) Immunohistochemical staining and Q) quantitative analysis of Col2 + chondrocytes per tissue area (mm 2 ) in Achilles tendons. Scale bar: 100 µm. R) TUNEL staining and S) quantitative analysis of apoptotic cells in Achilles tendons. Scale bar: 100 µm. T) Immunofluorescent staining of CD31 + cells and V) quantification of the fold change of blood vessels in Scx‐creERT2::Tgfbr2 f/f mice normalized to that of Tgfbr2 f/f mice in the Achilles tendons. Scale bar: 100 µm. T) Quantitative analysis of maximum tensile force and stiffness of Achilles tendons of Scx‐creERT2::Tgfbr2 f/f mice and their Tgfbr2 f/f littermates 8 weeks after DI. All data are shown as the mean ± standard deviation ( n = 8 mice per group). * p < 0.05 compared with Tgfbr2 f/f mice as determined by unpaired, 2‐tailed Student's t ‐test.

Article Snippet: Sections were processed for TUNEL staining according to the manufacturer's protocol (Promega, Madison, WI, catalog no. G3250, for Figures , , , and ; Promega, catalog no. G7360, for Figure ).

Techniques: Inhibition, Control, Immunohistochemical staining, Staining, TUNEL Assay, Standard Deviation, Knock-Out

Journal: Advanced Science

Article Title: Inhibition of Integrin α v β 6 Activation of TGF‐ β Attenuates Tendinopathy

doi: 10.1002/advs.202104469

Figure Lengend Snippet: Inducible knockout of αv in Scx + cells attenuates tendinopathy. A) Active TGF‐ β 1 concentration of tendon sections from Scx‐creERT2:: αv f/f and their WT αv f/f littermates upon tamoxifen injection. B) Immunostaining and C) quantification of pSmad2 + cells in tendons. Scale bar: 50 µm. D) SOFG staining of tendons in WT or Scx‐creERT2:: αv f/f mice. Proteoglycan (red) and collagenous matrix (green). Scale bar: 50 µm. E) Frequency distribution of tendon fiber orientation. F) Immunohistochemical staining and G) quantitative analysis of Col2 + chondrocytes per tissue area (mm 2 ) in Achilles tendons. Scale bar: 100 µm. H) TUNEL staining and I) quantitative analysis of apoptotic cells in tendons. Scale bar: 100 µm. J) Immunofluorescent staining of CD31 + cells and K) quantification of the fold change of blood vessels in Scx‐creERT2:: αv f/f mice normalized to that of WT mice in the Achilles tendons. Scale bar: 100 µm. Quantitative analysis of L) maximum tensile force and stiffness of tendons. All data are shown as the mean ± standard deviation ( n = 8 mice per group). * p < 0.05 compared to αv f/f mice as determined by unpaired, 2‐tailed Student's t ‐test.

Article Snippet: Sections were processed for TUNEL staining according to the manufacturer's protocol (Promega, Madison, WI, catalog no. G3250, for Figures , , , and ; Promega, catalog no. G7360, for Figure ).

Techniques: Knock-Out, Concentration Assay, Injection, Immunostaining, Staining, Immunohistochemical staining, TUNEL Assay, Standard Deviation