rabbit polyclonal anti gp91phox  (Bioss)

Journal: BMC Cardiovascular Disorders

Article Title: Aggravated endothelial endocrine dysfunction and intimal thickening of renal artery in high-fat diet-induced obese pigs following renal denervation

doi: 10.1186/s12872-020-01472-7

Figure Lengend Snippet: Effects of RDN treatment on NOX2 and 4-HNE expression. a NOX2, ( b ) 4-HNE. Data are expressed as the mean ± standard deviation. * p < 0.05, ** p < 0.01 vs. sham group; n = 5 per group. Abbreviations: RDN, renal denervation; NOX2, NADPH oxidase 2; 4-HNE,4-hydroxynonenal; GAPDH, glyceraldehyde phosphate dehydrogenase

Article Snippet: Protein samples were separated by SDS-PAGE (10% gel), transferred to PDVF membranes and blocked with 5% skim milk dissolved in 0.5% TBST for 1 h. Then, the membranes were incubated with the primary antibodies rabbit polyclonal anti-NADPH oxidase 2 (NOX2; 1:1000; bs-3889R, Bioss, Beijing, China), rabbit polyclonal anti-4-hydroxynonenal (4-HNE; 1:1000; ab46545, Abcam, Cambridge, UK), mouse monoclonal anti-endothelin 1 (ET-1; 1:500; abx100923, Abbexa, Cambridge, UK), rabbit polyclonal anti-endothelin A receptor (ET A R; 1:1000; ab117521, Abcam, Cambridge, UK), rabbit polyclonal anti-endothelin B receptor (ET B R; 1:1000; ab117529, Abcam, Cambridge, UK), rabbit polyclonal anti-endothelin converting enzyme 1 (ECE1; 1:1000; bs-1190R, Bioss Beijing, China), rabbit polyclonal anti-adenosine 5′-monophosphate (AMP)-activated protein kinase alpha 1/2 (AMPK; 1:1000; abx008836, Abbexa, Cambridge, UK), rabbit polyclonal anti-phosphorylated AMPK alpha (Thr172) (1:1000; 2531, CST, Boston, USA), rabbit polyclonal anti-endothelial nitric oxide synthase (eNOS; 1:1000; ab5589, Abcam, Cambridge, UK), rabbit polyclonal anti-phosphorylated eNOS (Ser1177) (1:1000; 9571, CST, Boston, USA), rabbit polyclonal anti-protein kinase B (Akt; 1:1000; 9272, CST, Boston, USA), rabbit polyclonal anti-phosphorylated Akt (Ser473) (1:1000; 9271, CST, Boston, USA), mouse monoclonal anti-glyceraldehyde phosphate dehydrogenase (GAPDH; 1:25000; GB13002-m-1, Servicebio, Wuhan, China), mouse monoclonal anti-histone H3 (11,000; GB13102–1, Servicebio, Wuhan, China), rabbit monoclonal anti-phosphorylated-I kappa B alpha (Ser32) (p-IκB alpha;1:1000;2859, CST, Boston, USA) and rabbit polyclonal anti- phosphorylated NF-kappa B p65 (Ser529) (p-NF-κB p65; 1:1000; LS-B652–50, LSBio, Seattle, USA) overnight at 4 °C.

Techniques: Expressing, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Aldosterone promotes renal interstitial fibrosis via the AIF-1/AKT/mTOR signaling pathway

doi: 10.3892/mmr.2019.10680

Figure Lengend Snippet: Effect of aldosterone on the protein expression levels of (A) AIF-1, p-PI3K, p-AKT, p-mTOR, NOX2 and Nrf2 in normal rats using western blotting. Aldosterone (0.75 µg/h subcutaneous infusion) increased the expression of (B) AIF-1, (C) p-PI3K, (D) p-AKT, (E) p-mTOR and (F) NOX2 and decreased the expression of (G) Nrf2 in normal rats, which was attenuated by spironolactone (100 mg/kg/day). *P<0.05 vs. con group, **P<0.05 vs. ald group, ***P<0.05 vs. ald + spir group. The data are presented as the mean ± standard deviation (n=3). AIF-1, allograft inflammatory factor-1; p-PI3K, phosphorylated phosphatidylinositol 3-kinase; p-AKT, phosphorylated AKT serine/threonine kinase; p-mTOR, phosphorylated mammalian target of rapamycin; NOX2, NADPH oxidase 2; Nrf2, nuclear transcription factor erythroid-related factor 2; con, control; ald, aldosterone; spir, spironolactone.

Article Snippet: The membranes were then incubated overnight at 4°C with the following primary antibodies: Rabbit anti-rat polyclonal AIF-1 (1:150; cat. no. ab153696; Abcam), rabbit anti-rat polyclonal Nrf2 antibody (1:200; cat. no. ab31163, Abcam), rabbit anti-rat polyclonal NOX2 antibody (1:300; cat. no. bs-3889R; Bioss Antibodies), rabbit anti-rat monoclonal p-PI3K p85 (1:200; cat. no. 4257; Cell Signaling Technology, Inc.), rabbit anti-rat monoclonal p-AKT (Ser473) (1:200; cat. no. 4060; Cell Signaling Technology, Inc.), rabbit anti-rat monoclonal p-mTOR (Ser2448) antibody (1:200; cat. no. #5536; Cell Signaling Technology, Inc.) and anti-β-actin monoclonal antibody (1:2,000; cat. no. A00702; GenScript).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Aldosterone promotes renal interstitial fibrosis via the AIF-1/AKT/mTOR signaling pathway

doi: 10.3892/mmr.2019.10680

Figure Lengend Snippet: (A) Western blot analysis of the protein expression levels of p-PI3K, p-AKT, p-mTOR, NOX2 and Nrf2 in the rat kidney. Aldosterone (0.75 µg/h subcutaneous infusion) upregulated the expression levels of (B) p-PI3K, (C) p-AKT, (D) p-mTOR and (E) NOX2 and downregulated the expression of (F) Nrf2, which was attenuated by spironolactone (100 mg/kg/day). *P<0.05 vs. con group, **P<0.05 vs. UUO group, ***P<0.05 vs. UUO + ald group. Data are presented as the mean ± standard deviation (n=3). p-PI3K, phosphorylated phosphatidylinositol 3-kinase; p-AKT, phosphorylated AKT serine/threonine kinase; p-mTOR; phosphorylated mammalian target of rapamycin; UUO, unilateral ureteric obstruction; NOX2, NADPH oxidase 2; Nrf2, nuclear transcription factor erythroid-related factor 2; con, control; ald, aldosterone; spir, spironolactone.

Article Snippet: The membranes were then incubated overnight at 4°C with the following primary antibodies: Rabbit anti-rat polyclonal AIF-1 (1:150; cat. no. ab153696; Abcam), rabbit anti-rat polyclonal Nrf2 antibody (1:200; cat. no. ab31163, Abcam), rabbit anti-rat polyclonal NOX2 antibody (1:300; cat. no. bs-3889R; Bioss Antibodies), rabbit anti-rat monoclonal p-PI3K p85 (1:200; cat. no. 4257; Cell Signaling Technology, Inc.), rabbit anti-rat monoclonal p-AKT (Ser473) (1:200; cat. no. 4060; Cell Signaling Technology, Inc.), rabbit anti-rat monoclonal p-mTOR (Ser2448) antibody (1:200; cat. no. #5536; Cell Signaling Technology, Inc.) and anti-β-actin monoclonal antibody (1:2,000; cat. no. A00702; GenScript).

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Aldosterone promotes renal interstitial fibrosis via the AIF-1/AKT/mTOR signaling pathway

doi: 10.3892/mmr.2019.10680

Figure Lengend Snippet: Effect of aldosterone on the protein expression levels of p-PI3K, p-AKT, p-mTOR, NOX2 and Nrf2 in RAW264.7 cells. Aldosterone (10-6 M) increased the expression levels of p-AKT, p-mTOR, NOX2 and Nrf2 via AIF-1 in RAW264.7 cells. (A) Western blot image of p-PI3K, p-AKT, p-mTOR, NOX2 and Nrf2 in different cells. Western blot analysis of (B) p-PI3K, (C) p-AKT, (D) p-mTOR, (E) NOX2 and (F) Nrf2. *P<0.05 vs. control, **P<0.05 vs. ald + pShuttle. Data are presented as the mean ± standard deviation (n=3). p-PI3K, phosphorylated phosphatidylinositol 3-kinase; p-AKT, phosphorylated AKT serine/threonine kinase; p-mTOR, phosphorylated mammalian target of rapamycin; NOX2, NADPH oxidase 2; Nrf2, nuclear transcription factor erythroid-related factor 2; AIF-1, allograft inflammatory factor-1; siRNA, small interfering RNA; over-exp, overexpression.

Article Snippet: The membranes were then incubated overnight at 4°C with the following primary antibodies: Rabbit anti-rat polyclonal AIF-1 (1:150; cat. no. ab153696; Abcam), rabbit anti-rat polyclonal Nrf2 antibody (1:200; cat. no. ab31163, Abcam), rabbit anti-rat polyclonal NOX2 antibody (1:300; cat. no. bs-3889R; Bioss Antibodies), rabbit anti-rat monoclonal p-PI3K p85 (1:200; cat. no. 4257; Cell Signaling Technology, Inc.), rabbit anti-rat monoclonal p-AKT (Ser473) (1:200; cat. no. 4060; Cell Signaling Technology, Inc.), rabbit anti-rat monoclonal p-mTOR (Ser2448) antibody (1:200; cat. no. #5536; Cell Signaling Technology, Inc.) and anti-β-actin monoclonal antibody (1:2,000; cat. no. A00702; GenScript).

Techniques: Expressing, Western Blot, Standard Deviation, Small Interfering RNA, Over Expression