Immunofluorescence Staining of HCV Core

Immunofluorescence staining of HCV core protein in OR6 cells and JFH1-infected Huh7.5.1 cells were performed as previously described [38 (link)]. OR6, Huh7.5.1 or JFH1 cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% TritonX-100, and blocked with 3% bovine serum albumin in PBS. The primary antibody was mouse anti-HCV core (Thermoscientific). The secondary antibody was goat anti–mouse Alexa Fluor 488 (Invitrogen). DAPI was added to the staining to monitor the nuclear structure. Fluorescence signals were observed by fluorescence microscopy (Zeiss, Axcio Observer A1).

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Tsai W.L., Chang T.H., Sun W.C., Chan H.H., Wu C.C., Hsu P.I., Cheng J.S, & Yu M.L. (2017). Metformin activates type I interferon signaling against HCV via activation of adenosine monophosphate-activated protein kinase. Oncotarget, 8(54), 91928-91937.

Publication 2017

mouse anti-HCV core goat anti–mouse Alexa Fluor 488 Axcio Observer A1

Alexa fluor 488 Anti hcv Antibody Bovine serum albumin Cells Dapi Fluorescence Fluorescence microscopy Goat Hcv core protein Immunofluorescence Mouse Paraformaldehyde

Corresponding Organization :

Other organizations : National Yang Ming Chiao Tung University, Kaohsiung Veterans General Hospital, Chung Hwa University of Medical Technology, Kaohsiung Medical University, Kaohsiung Medical University Chung-Ho Memorial Hospital, National Sun Yat-sen University

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Variable analysis

independent variables

Immunofluorescence staining of HCV core protein

dependent variables

Fluorescence signals

control variables

Cell lines: OR6, Huh7.5.1, and JFH1
Fixation with 4% paraformaldehyde
Permeabilization with 0.5% TritonX-100
Blocking with 3% bovine serum albumin in PBS
Primary antibody: mouse anti-HCV core (Thermoscientific)
Secondary antibody: goat anti–mouse Alexa Fluor 488 (Invitrogen)
DAPI staining to monitor nuclear structure

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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