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. 2015 Mar 4;89(10):5462–5477. doi: 10.1128/JVI.03197-14

FIG 9.

FIG 9

Interaction of IKK-α with HCV nonstructural proteins. (A and B) Colocalization of IKK-α with HCV NS3 and NS5B proteins. Huh-7.5.1 cells were infected with JFH-1 HCVcc for 48 h (A) or were transfected with various plasmids encoding HCV proteins for 24 h (B). The subcellular localization of IKK-α and its association with HCV proteins were detected and analyzed by immunofluorescence and confocal microscopy. Scale bars, 20 μm. Pearson's correlation coefficient (CR) was determined on the selected areas (in white boxes). (C and D) Cells were transfected with plasmids encoding various Flag-tagged HCV proteins and/or HA-IKK-α for 24 h. Immunoprecipitation (IP) with anti-HA, anti-Flag, or IgG control antibody was performed, followed by Western blotting (WB) using anti-Flag or anti-HA antibody. (E) Cells were transfected with in vitro-transcribed EGFP control RNA (Ctrl), SGR2a, or JFH-1 RNA for 48 h. Immunoprecipitation on whole-cell lysate (WCL) with anti-IKK-α or anti-DDX3X antibody was conducted, followed by Western blotting using anti-NS3 antibody. HC, IgG heavy chain.