Journal: The Journal of biological chemistry

Article Title: Analysis of small latent transforming growth factor-beta complex formation and dissociation by surface plasmon resonance. Absence of direct interaction with thrombospondins.

doi: 10.1074/jbc.272.26.16329

Figure Lengend Snippet: FIG. 1. LAP forms a complex with immobilized TGFb1. TGFb1 was immobilized (around 600 RU), and LAP was then injected to form small latent TGFb1 complex. A, sensorgrams (relative response in RU after background subtraction versus time in sec) of LAP binding (bottom to top curve: 50, 125, 150, 175, and 200 nM of LAP). B, plots of ks versus LAP concentration.

Article Snippet: Ultrapure human TGFb1 was from Genzyme (Cambridge, MA), recombinant human LAP (TGFb1), and recombinant latent TGFb1 were from R&D Systems (Abingdon, UK).

Techniques: Injection, Binding Assay, Concentration Assay

Journal: The Journal of biological chemistry

Article Title: Analysis of small latent transforming growth factor-beta complex formation and dissociation by surface plasmon resonance. Absence of direct interaction with thrombospondins.

doi: 10.1074/jbc.272.26.16329

Figure Lengend Snippet: FIG. 2. TGFb1 forms a complex with immobilized LAP. LAP was immobilized (around 800 RU) and then TGFb1 was injected to form small latent TGFb1 complex. Sensorgrams (relative response in RU after background subtraction versus time in sec) of TGFb1 binding (bottom to top curve: 50, 80, and 100 nM of TGFb1).

Article Snippet: Ultrapure human TGFb1 was from Genzyme (Cambridge, MA), recombinant human LAP (TGFb1), and recombinant latent TGFb1 were from R&D Systems (Abingdon, UK).

Techniques: Injection, Binding Assay

Journal: The Journal of biological chemistry

Article Title: Analysis of small latent transforming growth factor-beta complex formation and dissociation by surface plasmon resonance. Absence of direct interaction with thrombospondins.

doi: 10.1074/jbc.272.26.16329

Figure Lengend Snippet: FIG. 3. Activation of latent TGFb1 complex. TGFb1 was immo- bilized (around 600 RU), and then LAP (50 nM) was injected to form small latent TGFb1 complex. A, over this complex, 25 ml of 6 M guani- dine chloride (GnCl) was injected at a flow of 5 ml/min, followed by another injection of LAP (50 nM) and then a second injection of 6 M GnCl. B, over this complex, bottom to top curve: 50 ml of HBS (CTL), pH 8.6; *, plasmin (0.1 units/ml in HBS, pH 8.6); or **, heat-inactivated plasmin (0.1 units/ml in HBS, pH 8.6) was injected at a flow of 2 ml/min.

Article Snippet: Ultrapure human TGFb1 was from Genzyme (Cambridge, MA), recombinant human LAP (TGFb1), and recombinant latent TGFb1 were from R&D Systems (Abingdon, UK).

Techniques: Activation Assay, Injection

Journal: The Journal of biological chemistry

Article Title: Analysis of small latent transforming growth factor-beta complex formation and dissociation by surface plasmon resonance. Absence of direct interaction with thrombospondins.

doi: 10.1074/jbc.272.26.16329

Figure Lengend Snippet: FIG. 4. Absence of association of TSP1 and TSP2 with LAP, TGFb1, or small latent TGFb1 forms. A, TSP1 (50 nM) was injected over immobilized LAP, TGFb1, and a control (CTL) immobilized flow cell. B, TSP2 (50 nM) was injected over immobilized LAP, TGFb1, and a control immobilized flow cell. C, LAP was immobilized and then TGFb1 (50 nM) was injected to form small latent TGFb1 complex, and over this complex, TSP2 (50 nM) or TSP1 (50 nM) was injected. D, vice versa of C. TGFb1 was immobilized and then LAP (50 nM) was injected to form small latent TGFb1 complex, and over this complex, TSP1 (50 nM) or TSP2 (50 nM) was injected.

Article Snippet: Ultrapure human TGFb1 was from Genzyme (Cambridge, MA), recombinant human LAP (TGFb1), and recombinant latent TGFb1 were from R&D Systems (Abingdon, UK).

Techniques: Injection, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: MicroRNA-150 relieves vascular remodeling and fibrosis in hypoxia-induced pulmonary hypertension.

doi: 10.1016/j.biopha.2018.11.058

Figure Lengend Snippet: Fig. 4. Effect of miR-150 on pulmonary fibrosis of pulmonary hypertension rats. (A) Pulmonary fibrosis was detected by Masson’s staining (200× magnification). Scale bars, 100 μm. (B) The area of pulmonary fibrosis was calculated and shown. The mRNA expressions of TGF-β1 (C) and collagen I (D) in lung tissues were evaluated by qPCR. (E) The protein levels of TGF-β1 and collagen I in lung tissues were measured by western blot assay. (F)&(G) Relative grey values of the protein bands were shown. (H) The expressions of TGF-β1 and collagen I in lung tissues were detected by immunohistochemical staining (400× magnification). Scale bars, 50 μm. Data were presented as mean ± SD. **P < 0.01, ***P < 0.001 versus the indicated group.

Article Snippet: The membranes were blocked in 5% nonfat milk for 1 h at room temperature, then incubated with primary antibodies against TGF-β1 (1:400, BOSTER, China), Collagen I (1:300, BOSTER, China), p-AKTser473 (1:500, KeyGen, China), AKT (1:500, KeyGen, China), p-mTORser2481 (1:500, Sangon Biotech, China), mTOR (1:1000, Cell signaling Technology, USA), and β-actin (1:500, Bioss, China) at 4°C overnight, followed by incubation with HRP-labeled Goat AntiRabbit or Goat Anti-Mouse IgG (1:5000, Beyotime, China) at 37°C for 45min.

Techniques: Staining, Western Blot, Immunohistochemical staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: MicroRNA-150 relieves vascular remodeling and fibrosis in hypoxia-induced pulmonary hypertension.

doi: 10.1016/j.biopha.2018.11.058

Figure Lengend Snippet: Fig. 6. Effect of miR-150 on the expressions of fibrosis-related molecules. The mRNA expressions of TGF-β1 (A) and collagen I (B) in PASMCs were measured by qPCR. (C) The protein levels of TGF-β1 and collagen I in PASMCs were detected by western blot assay. (D)&(E) Relative grey values of the protein bands were shown. Data were presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 versus the indicated group.

Article Snippet: The membranes were blocked in 5% nonfat milk for 1 h at room temperature, then incubated with primary antibodies against TGF-β1 (1:400, BOSTER, China), Collagen I (1:300, BOSTER, China), p-AKTser473 (1:500, KeyGen, China), AKT (1:500, KeyGen, China), p-mTORser2481 (1:500, Sangon Biotech, China), mTOR (1:1000, Cell signaling Technology, USA), and β-actin (1:500, Bioss, China) at 4°C overnight, followed by incubation with HRP-labeled Goat AntiRabbit or Goat Anti-Mouse IgG (1:5000, Beyotime, China) at 37°C for 45min.

Techniques: Western Blot