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Image Search Results
Journal: Acta Pharmaceutica Sinica. B
Article Title: Up-regulation of glycolipid transfer protein by bicyclol causes spontaneous restriction of hepatitis C virus replication
doi: 10.1016/j.apsb.2019.01.013
Figure Lengend Snippet: GLTP restricts HCV replication by interrupting the interaction between VAP-A and NS5A. (A) The GLTP directly bound VAP-A in a co-IP assay. The protein VAP-A was as the bait protein, and the binding was confirmed by detecting His-tag of the GLTP-His. (B) The level of GLTP negatively affected the level of NS5A bound to VAP-A in the co-IP assay. The protein VAP-A was as the bait protein, and the Huh7.5 cells transfected with expression plasmid or shRNA plasmid for GLTP were used. (C) GLTP had no direct interaction with NS5A in the co-IP assay. The protein GLTP served as the bait protein. (D) Changing the level of GLTP did not influence the level of intracellular VAP-A protein. The Huh7.5 cells were transfected with the expression plasmid or shRNA plasmid for GLTP; intracellular proteins were detected in 72 h by WB ( n = 3; ** P < 0.01 vs. control). The anti-HCV activity and a co-IP assay for mutants of GLTP. The Huh7.5 cells were transfected with GLTP of wild type or W site-directed mutant (E) or amino acid 33-35 mutant (F), and infected with HCV (45 IU/cell). After 72 h incubation, intracellular proteins were extracted and detected with WB (left, n = 3; * P < 0.05 and ** P < 0.01 vs. control). For co-IP assay, the protein VAP-A was as the bait protein (right). The detail method of the co-IP assay was in the Material and Methods. The experiments were carried out for over 3 times. The protein bands presented in the figure showed the results of a representative experiment. The data presented are the mean ± standard deviation of three independent experiments and the Student׳s t -test was used in A–F.
Article Snippet: Antibodies for beta-Actin (3700s) and
Techniques: Co-Immunoprecipitation Assay, Binding Assay, Transfection, Expressing, Plasmid Preparation, shRNA, Control, Activity Assay, Mutagenesis, Infection, Incubation, Standard Deviation