Journal: Yonsei Medical Journal

Article Title: Changes in Inward Rectifier K + Channels in Hepatic Stellate Cells During Primary Culture

doi: 10.3349/ymj.2008.49.3.459

Figure Lengend Snippet: Relative inward rectifier K + channel and SUR gene expression in HSC. (A) The inward rectifier K + channel α-subunit gene expression in HSC was measured using real-time RT-PCR. The bar graphs show the relative gene expression for each inward rectifier K + channel subfamily (K ir 1.1, K ir 2.1 - K ir 2.4, K ir 3.1 - K ir 3.4, K ir 4.1 - K ir 4.2, K ir 5.1, K ir 6.1 - K ir 6.2, and K ir 7.1). (B) SUR gene expression was measured. The bar graphs show the relative gene expression for SUR1, SUR2A, and SUR2B. The HSC were used at 1 day, 1 week, 2 weeks, and 3 weeks of culture. The expression levels were normalized to GAPDH and calibrated by K ir 2.1 expression in the HSC cultured for 1 day. The data are shown as the mean ± SEM (n = 3). (C) K ir 2.1 and K ir 6.1 protein expression were measured by Western blotting. There was a band (55 kD) in the anti-K ir 2.1 membrane (Upper) as well as the anti-K ir 6.1 membrane (Lower). Both membranes were re-probed with anti-GAPDH antibody as shown. The data is representative of three independent experiments.

Article Snippet: The anti-K ir 2.1 and anti-K ir 6.1 antibodies were obtained from Alomone Labs (Jerusalem, Israel), the anti-GAPDH antibody was acquired from Novus Biologicals (Littleton, CO, USA), and Western Lightning was supplied by PerkinElmer Life Sciences (Gaithersburg, MD, USA).

Techniques: Gene Expression, Quantitative RT-PCR, Expressing, Cell Culture, Western Blot, Membrane

Journal: Experimental and Therapeutic Medicine

Article Title: Compound Astragalus and Salvia miltiorrhiza extract inhibits hepatocarcinogenesis via modulating TGF-β/TβR and Imp7/8

doi: 10.3892/etm.2018.6292

Figure Lengend Snippet: CASE decreases the protein expression of TGF-β 1 , TβRI and TβRII. The effects of CASE on TGF-β 1 , TβRI and TβRII expression in rats treated with DEN were assessed by western blotting (A-C) 12 and (D-F) 16 weeks after the induction of hepatocellular carcinoma by DEN. The proteins were extracted from frozen liver tissues. TGF-β 1 , TβRI and TβRII proteins were analyzed using anti-TGF-β 1 , -TβRI, -TβRII and -GAPDH antibodies. Intensities of TGF-β 1 , TβRI and TβRII bands were normalized to those of GAPDH in the corresponding treatment groups. The ratios of the TGF-β 1 , TβRI or TβRII protein to GAPDH in the normal groups were assigned a value of 1. Data are expressed as mean ± standard deviation (n=3). ## P<0.01 vs. the control group. **P<0.01 vs. the DEN group. DEN, diethylinitrosamine; CASE, Compound Astragalus and Salvia miltiorrhiza extract; TGF-β 1 , transforming growth factor β-1; TβR, transforming growth factor-β receptor.

Article Snippet: The primary antibodies utilized were as follows: Rabbit anti-TGF-β 1 Ab (cat. no. sc-146; 1:800; Santa Cruz Biotechnology, Inc.), rabbit anti-TβRI Ab (cat. no. sc-398; 1:800; Santa Cruz Biotechnology, Inc.) and goat anti-TβRII Ab (cat. no. sc-33929; 1:800; Santa Cruz Biotechnology, Inc.), mouse anti-GAPDH Ab (cat. no. TA-08; dilution, 1:5,000; Origene Technologies, Inc., Rockville, MD, USA), rabbit anti-GST-P1 Ab (cat. no. SAB3500265; 1:1,000; Sigma-Aldrich, Merck KGaA), rabbit anti-Imp7 Ab (cat. no. ab99273; 1:5,000; Abcam, Cambridge, UK) and rabbit anti-Imp8 Ab (cat. no. ab72109; 1:5,000; Abcam).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Compound Astragalus and Salvia miltiorrhiza extract inhibits hepatocarcinogenesis via modulating TGF-β/TβR and Imp7/8

doi: 10.3892/etm.2018.6292

Figure Lengend Snippet: High and medium CASE doses decrease TβRI and TβRII protein expression in MFBs and HepG2 cells. Cells were starved for 24 h in serum-free medium and in the absence or presence of CASE (20, 40 or 80 µg/ml), subsequently treated with TGF-β 1 . Total proteins were extracted from (A) rat myofibroblasts and (B) HepG2 cells. The expression of TβRI and TβRII proteins were analyzed by western blotting using anti-TβRI, -TβRII and -GAPDH antibodies. Intensities of TβRI and TβRII bands were normalized to those of GAPDH in the corresponding treatment groups. The ratios of the TβRI and TβRII protein to GAPDH without exogenous TGF-β 1 were assigned a value of 1. Data are expressed as mean ± standard deviation (n=3). ## P<0.01 vs. the control group. *P<0.05 and **P<0.01 vs. the TGF-β 1 group. CASE, Compound Astragalus and Salvia miltiorrhiza extract; TGF-β 1 , transforming growth factor β-1; TβR, transforming growth factor-β receptor.

Article Snippet: The primary antibodies utilized were as follows: Rabbit anti-TGF-β 1 Ab (cat. no. sc-146; 1:800; Santa Cruz Biotechnology, Inc.), rabbit anti-TβRI Ab (cat. no. sc-398; 1:800; Santa Cruz Biotechnology, Inc.) and goat anti-TβRII Ab (cat. no. sc-33929; 1:800; Santa Cruz Biotechnology, Inc.), mouse anti-GAPDH Ab (cat. no. TA-08; dilution, 1:5,000; Origene Technologies, Inc., Rockville, MD, USA), rabbit anti-GST-P1 Ab (cat. no. SAB3500265; 1:1,000; Sigma-Aldrich, Merck KGaA), rabbit anti-Imp7 Ab (cat. no. ab99273; 1:5,000; Abcam, Cambridge, UK) and rabbit anti-Imp8 Ab (cat. no. ab72109; 1:5,000; Abcam).

Techniques: Expressing, Western Blot, Standard Deviation