rabbit polyclonal anti-cav-1  (Santa Cruz Biotechnology)

Journal: Pulmonary Circulation

Article Title: Hypoxia‐induced pulmonary hypertension upregulates eNOS and TGF‐β contributing to sex‐linked differences in BMPR2 +/R899X mutant mice

doi: 10.1002/pul2.12163

Figure Lengend Snippet: Depletion of pulmonary BMPRII‐short and long fragments in humanized BMPR2 +/R899X mutant mice caused vascular remodeling independent of pulmonary caveolin‐1 expression under normoxic conditions. Male and female wild type (WT) and BMPR2 +/R899X mice were genotyped using amplification refractory mutation system PCR (ARMS‐PCR) with 428 bp indicating the full‐length BMPR2 and 193 bp, amplification of the BMPR2 mutated fragment. Genotyped animals were kept under normoxia for 1 month, and then perfused lung tissue was obtained to evaluate the expression level of BMPRII short and long fragment (SF/LF), phosphorylated‐SMAD1/5/8 (P‐SMAD1/5/8), β‐actin, and caveolin‐1 (Cav‐1) (a, b) by western blot or fixed for histological analysis of the microvessel area and collagen deposition using Picrosirius FAST Red staining (c, d). The percentage of fully muscularized microvessels was quantified using von Willebrand Factor (vWF; Red) and alpha‐smooth muscle actin (α‐SMA; green) staining (b, d). Normally distributed data were evaluated using the Student t ‐test. * p < 0.05, ** p < 0.01, **** p < 0.001, ns, non significant.

Article Snippet: Mouse monoclonal anti‐eNOS, anti‐β‐actin and rabbit polyclonal anti‐Cav‐1 were purchased from BD PharMingen.

Techniques: Mutagenesis, Expressing, Amplification, Western Blot, Staining

Journal: Pulmonary Circulation

Article Title: Hypoxia‐induced pulmonary hypertension upregulates eNOS and TGF‐β contributing to sex‐linked differences in BMPR2 +/R899X mutant mice

doi: 10.1002/pul2.12163

Figure Lengend Snippet: Chronic hypoxia exposure increased the number of muscularized microvessels within the lungs of female BMPR2 +/R899X mutant mice. (a, b) Mouse lung sections from male (light blue bars and circles) and female (light red bars and circles) BMPR2 +/R899X mice (normoxia and hypoxia) were stained with an antibody against smooth muscle actin (α‐SMA; red), and the nuclei stained using DAPI (blue). Then, the number of fully muscularized α‐SMA positive pulmonary microvessels with diameters smaller than 100 μm (scale bar: 50 μm) were quantified per lung tissue area (mm 2 ; four areas per animal) using confocal microscopy. (c–f) Mouse lung tissue from all groups was used to quantify the expression levels of the total and monomeric endothelial nitric oxide synthase (eNOS), total caveolin‐1 (Cav‐1), and endothelin‐1(ET‐1). β‐actin was used as loading control. Normally distributed data were evaluated using two‐way analysis of variance followed by Bonferroni post hoc test ( n = 4 mice per group). * p < 0.05, ** p < 0.01, ns, non significant.

Article Snippet: Mouse monoclonal anti‐eNOS, anti‐β‐actin and rabbit polyclonal anti‐Cav‐1 were purchased from BD PharMingen.

Techniques: Mutagenesis, Staining, Confocal Microscopy, Expressing