dab staining with nickel  (Vector Laboratories)

Journal: Redox Biology

Article Title: Novel and superior treatment of pulmonary hypertension with netrin-1 derived, modified and improved small peptides

doi: 10.1016/j.redox.2025.103710

Figure Lengend Snippet: Novel, modified peptides derived from netrin-1 exerted augmented effects in attenuating pulmonary vascular muscularization and cell proliferation in pulmonary hypertensive mice . Pulmonary hypertension was induced in mice by exposure to normobaric hypoxia (10% O 2 ) for three weeks. Osmotic pump was used to deliver modified peptides derived from netrin-1, V1P, V2P, V3P, V1S, V1T, V1D, V1C continuously. (A,D) Representative images and quantitative data of smooth muscle actin (SMA) expression by immunohistochemical staining using DAB substrate. (B,E) Representative images and quantitative data of proliferating cell nuclear antigen (PCNA) expression by immunohistochemical staining using DAB substrate. (C,F) Representative images and quantitative data of collagen deposition by Masson's Trichrome staining. Data indicate that pulmonary vascular muscularization, cell proliferation and fibrosis in hypoxia induced pulmonary hypertensive mice were completely attenuated by treatment with the modified netrin-1 peptides. Data are shown as Mean ± SEM. ∗∗∗p < 0.001 vs. Normoxia; ###p < 0.001 vs. Hypoxia. Normoxia (n = 5), Hypoxia (n = 5), Hypoxia + V1P (n = 5), Hypoxia + V2P (n = 5), Hypoxia + V3P (n = 5), Hypoxia + V1S (n = 5), Hypoxia + V1T (n = 5), Hypoxia + V1D (n = 5), Hypoxia + V1C (n = 5).

Article Snippet: For DAB staining, sections were incubated with secondary antibodies (1:200, Cat# PK-6102-mouse, NC9293436-rabbit, Vecta stain ABC kit, Vector labs, Burlingame, CA, USA) and stained using a 3,3-diaminobenzidine (DAB) substrate according to the manufacturer's instructions (MilliporeSigma, Cat# D3939, St. Louis, MO, USA).

Techniques: Modification, Derivative Assay, Expressing, Immunohistochemical staining, Staining