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: International journal of oncology

Article Title: Expression of ribophorine II is a promising prognostic factor in human gastric adenocarcinoma.

doi: 10.3892/ijo.2016.3822

Figure Lengend Snippet: Figure 4. Detection of genome editing at RPN2 gene. (A) Cloning target RPN2 sequence using the Guide-it CRISPR/Cas9 system. A red line under AGG indicates protospacer-adjacent motif. (B) Fluorescence analysis shows that transfection with RPN2-Cas9-GFP plasmid increased GFP-expressing cells in MKN74 and KATO III cell lines. Fluorescence cell imaging were visualized 96 h following transfection. (C) Indel frequency was measured using the T7E1 assay and the percentages are shown at the bottom of each lane. (D) Protein extracted from fluorescence positive cells was analyzed for RPN2 and GAPDH by western blot assay. (E) Sanger sequencing of PCR products around gRNA binding site (red letters) and protospacer adjacent motif (PAM) site (blue letters). Wild-type reference sequences are given on the top.

Article Snippet: Anti-RPN2 Ab was from Aviva Systems and anti-GAPDH Ab was from IMGENEX.

Techniques: Cloning, Sequencing, CRISPR, Fluorescence, Transfection, Plasmid Preparation, Expressing, Imaging, Western Blot, Binding Assay

Journal: International journal of oncology

Article Title: Expression of ribophorine II is a promising prognostic factor in human gastric adenocarcinoma.

doi: 10.3892/ijo.2016.3822

Figure Lengend Snippet: Figure 6. Relationship between RPN2 and p53 in gastric adenocarcinoma. (A) p53 protein stained in brown was detected in the nucleus of primary gastric adenocarcinoma lesions. (B) Evidence for knockdown of RPN2 caused by transfection with siRNA. MKN45 cells were transfected with scrambled siRNA (control); GAPDH was used as loading control. RPN2 siRNA followed by western blot assays using anti-RPN2 antibodies. (C) The knockout of RPN2 expression in MKN45 cell line did not reduce the ability of invasion. Data are expressed as mean values ± SD from triplicate experiments.

Article Snippet: Anti-RPN2 Ab was from Aviva Systems and anti-GAPDH Ab was from IMGENEX.

Techniques: Staining, Knockdown, Transfection, Control, Western Blot, Knock-Out, Expressing