Journal: Experimental and Therapeutic Medicine

Article Title: Fucoidan elevates surface organic cation transporter 2 expression via upregulation of protein kinase A in uric acid nephropathy

doi: 10.3892/etm.2017.5077

Figure Lengend Snippet: Fucoidan activates PKA and upregulates surface OCT2 in the renal tissue of a rat model of UAN. (A) Western blot analysis of the PKA 2β, p-PKA 2β, surface OCT2 and total OCT2 protein level in renal tissues. GAPDH was used as a loading control. (B) Densitometry plot of results from western blot analysis. Relative expression levels were normalized to GAPDH. Data are presented as the mean ± standard error of the mean (n=3). (C) Immunohistochemistry analysis of PKA expression in kidney tissues of rats with UAN. Magnification, ×400. *P<0.05; **P<0.01. PKA, protein kinase A; NC, normal control; UAN, uric acid nephropathy; OCT2, organic cation transporter 2; p, phosphorylated.

Article Snippet: The membranes were blocked with Tris-buffered saline-Tween-20 containing 2% bovine serum albumin (Sigma-Aldrich; Merck KGaA) at room temperature for 2 h, and the membranes were then incubated with specific antibodies against OCT2 (sc-233; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; 1:1,500), PKA 2β (sc-365615; Santa Cruz Biotechnology, Inc; 1:1,500), p-PKA 2β (sc-293036; Santa Cruz Biotechnology, Inc.; 1:1,000), GAPDH (ab8245; Abcam, Cambridge, UK; 1:8,000).

Techniques: Western Blot, Control, Expressing, Immunohistochemistry

Journal: Experimental and Therapeutic Medicine

Article Title: Fucoidan elevates surface organic cation transporter 2 expression via upregulation of protein kinase A in uric acid nephropathy

doi: 10.3892/etm.2017.5077

Figure Lengend Snippet: Surface OCT2 is upregulated by fucoidan in OCT2-expressing COS-7 cells. OCT2-expressing COS-7 cells were treated with fucoidan (500 µg/ml) or phosphate-buffered saline (NC) for 24 h. (A) Western blot analysis was performed to evaluate the protein expression of PKA 2β, p-PKA 2β, total OCT2 and surface OCT2. (B) Densitometry plot of results from western blot analysis. Relative expression levels were normalized to GAPDH. Data are expressed as the mean ± standard error of the mean (n=3). *P<0.05 vs. NC. PKA, protein kinase A; NC, normal control; OCT2, organic cation transporter 2; p, phosphorylated.

Article Snippet: The membranes were blocked with Tris-buffered saline-Tween-20 containing 2% bovine serum albumin (Sigma-Aldrich; Merck KGaA) at room temperature for 2 h, and the membranes were then incubated with specific antibodies against OCT2 (sc-233; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; 1:1,500), PKA 2β (sc-365615; Santa Cruz Biotechnology, Inc; 1:1,500), p-PKA 2β (sc-293036; Santa Cruz Biotechnology, Inc.; 1:1,000), GAPDH (ab8245; Abcam, Cambridge, UK; 1:8,000).

Techniques: Expressing, Saline, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Fucoidan elevates surface organic cation transporter 2 expression via upregulation of protein kinase A in uric acid nephropathy

doi: 10.3892/etm.2017.5077

Figure Lengend Snippet: PKA upregulates surface OCT2 in OCT2-expressing COS-7 cells. OCT2-expressing COS-7 cells were treated with PKA activator, 8-bromoadenosine 3′,5′-cyclic monophosphate (1 µmol/l) or phosphate-buffered saline (NC) for 12 h. (A) Western blot analysis was performed to evaluate the protein expression of surface OCT2, total OCT2 and intracellular OCT2. (B) Densitometry plot of results from western blot analysis. Relative expression levels were normalized to GAPDH. Data are presented as the mean ± standard error of the mean (n=3). *P<0.05 and **P<0.01 vs. NC. PKA, protein kinase A; NC, normal control; OCT2, organic cation transporter 2.

Article Snippet: The membranes were blocked with Tris-buffered saline-Tween-20 containing 2% bovine serum albumin (Sigma-Aldrich; Merck KGaA) at room temperature for 2 h, and the membranes were then incubated with specific antibodies against OCT2 (sc-233; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; 1:1,500), PKA 2β (sc-365615; Santa Cruz Biotechnology, Inc; 1:1,500), p-PKA 2β (sc-293036; Santa Cruz Biotechnology, Inc.; 1:1,000), GAPDH (ab8245; Abcam, Cambridge, UK; 1:8,000).

Techniques: Expressing, Saline, Western Blot, Control