rhgdf7  (R&D Systems)

Journal: Molecular Therapy

Article Title: TET-dependent GDF7 hypomethylation impairs aqueous humor outflow and serves as a potential therapeutic target in glaucoma

doi: 10.1016/j.ymthe.2020.12.030

Figure Lengend Snippet: Excessive GDF7 protein-promoted fibrosis in cultured TM cells (A) In cultured TM cells, the expression level of fibrosis-related markers was presented with immunofluorescence, and the collagen accumulation in TM cells was presented by Masson stain (n = 3 per group). (B) The mRNA levels of N-cadherin (N-cad), α-smooth muscle actin (α-SMA), and fibronectin (FN) were tested in recombinant human GDF7 (rhGDF7)-treated TM cells by quantitative real-time PCR assay (n = 3 per group). (C and D) As presented in western blot results, the protein levels of N-cad, α-SMA, and FN were measured in NTM cells with or without rhGDF7 treatment (n = 3 per group). Scale bars, 20 μm. The data represent the mean ± SD. Compared with NTM cells: ∗∗p < 0.01. Compared with rhGDF7 group: #p < 0.05, ##p < 0.01. nGDF7, GDF7 neutralizing antibody; Col I, collagen I.

Article Snippet: For each immunization, 100 mL of the immunogen mixture, containing 30–60 mg of filter-sterilized rhGDF7 and 60 mg of poly I:C in PBS, was injected into both inner rear thighs of mice.

Techniques: Cell Culture, Expressing, Immunofluorescence, Staining, Recombinant, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Therapy

Article Title: TET-dependent GDF7 hypomethylation impairs aqueous humor outflow and serves as a potential therapeutic target in glaucoma

doi: 10.1016/j.ymthe.2020.12.030

Figure Lengend Snippet: GDF7 promoted fibrosis through the BMPR2/Smad1, -5, and -9 pathway (A) Fibrosis was determined by immunofluorescence labeling with N-cad, α-SMA, and FN together with Masson staining in cultured TM cells. (B) The mRNA levels of GDF7 and fibrosis-related markers N-cad, α-SMA, and FN were tested by quantitative real-time PCR in four groups of TM cells (n = 3 per group). (C and D) The ratio between phosphorylated and total Smad1, -5, and -9 and Smad4 protein was measured and analyzed by immunoblot in TM cells (n = 3 per group). Scale bars, 20 μm. The data were represented as mean ± SD. Compared with rhGDF7 group: ∗p < 0.05, ∗∗p < 0.01. Compared with NTM cells: #p < 0.05.

Article Snippet: For each immunization, 100 mL of the immunogen mixture, containing 30–60 mg of filter-sterilized rhGDF7 and 60 mg of poly I:C in PBS, was injected into both inner rear thighs of mice.

Techniques: Immunofluorescence, Labeling, Staining, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Therapy

Article Title: TET-dependent GDF7 hypomethylation impairs aqueous humor outflow and serves as a potential therapeutic target in glaucoma

doi: 10.1016/j.ymthe.2020.12.030

Figure Lengend Snippet: GDF7 neutralization protected TM function in rhesus monkeys (A) Simplified workflow of animal experiment. In brief, rhGDF7 was delivered into the anterior chamber three times on the first days of weeks 0, 2, and 4. The nGDF7 was injected into the anterior chamber for three times on the first days of weeks 10, 12, and 14. Ophthalmic examinations were conducted every 2 weeks throughout the entire process. The TM samples were harvested on the end of week 28. (B) The aqueous humor (AH) outflow facility was measured throughout the whole experiment. (C) Curves presented the IOP change in response to rhGDF7 delivery or/and nGDF7 treatment in monkey eyes (n = 4 per group). (D) Immunofluorescence labeling with N-cad, α-SMA, and FN was conducted in monkey TM tissues, and collagen accumulation was determined by Masson trichrome staining (n = 4 per group). Scale bars, 20 μm.

Article Snippet: For each immunization, 100 mL of the immunogen mixture, containing 30–60 mg of filter-sterilized rhGDF7 and 60 mg of poly I:C in PBS, was injected into both inner rear thighs of mice.

Techniques: Neutralization, Injection, Immunofluorescence, Labeling, Staining