nb100-91995  (Bio-Techne corporation)

Journal: Scientific Reports

Article Title: P311 promotes renal fibrosis via TGFβ1/Smad signaling

doi: 10.1038/srep17032

Figure Lengend Snippet: (A) RNA was isolated in kidneys from P311 +/+ (n = 6) and P311 −/− (n = 6) mice 7 days after UUO or sham operation. TGF-β1 mRNA expression was determined by real-time PCR. (B) Representative photomicrographs of TGF-β1-specific immunohistochemical staining of UUO kidneys from P311 +/+ and P311 −/− mice on day 7. Black arrowhead indicates the TGF-β1-positive region. Bottom panel: negative controls for the immunohistochemical staining of TGF-β1 on the obstructed kidneys from both P311 +/+ and P311 −/− mice. ( C ) TGF-β1-positive region was quantified in stained sections from P311 +/+ (n = 6) and P311 −/− (n = 6) mice 7 days after UUO. ( D ) Western blot analysis of TGF-β1 protein levels. TGF-β1 protein levels in each treatment group (n = 3 per group) were quantified. A-D are representative of at least three similar experiments. Scale bar: 100μm. Data are presented as the mean ± SD. *P < 0.05; **P < 0.01.

Article Snippet: For immunohistochemical staining, sections were incubated with an antigen retrieval solution at 37 °C for 30 min. After antigen retrieval, sections were incubated with 3% H 2 O 2 for 20 min at room temperature, with goat serum at 37 °C for 30 min, and with P311 Ab (1:400 dilution; Bioss, China, bs-0427R), α-SMA Ab (1:200 dilution; Abcam, USA, ab5694), TGF-β1 Ab (1:400 dilution; Novus, USA, NB100-91995) or F4/80 Ab (1:100 dilution; Abcam, USA, ab111101) overnight at 4 °C.

Techniques: Isolation, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Western Blot