Journal: Emerging Microbes & Infections

Article Title: Erp57 facilitates ZIKV-induced DNA damage via NS2B/NS3 complex formation

doi: 10.1080/22221751.2024.2417864

Figure Lengend Snippet: ERp57 is critical for ZIKA nonstructural protein expression and function. (A) Representative immunofluorescent staining images show the localization of ZIKV NS5 (green). WT RD cells were seeded in 24-well plates with pre-coated cover glasses, then infected with ZIKA virus (MOI = 1) for the indicated time. Cells were fixed and stained with anti-NS5 (green) fluorescence antibody and DAPI (blue) and visualized under confocal microscope. (B) and (C) Both wild type RD cells and knockout cells were infected by ZIKV (MOI = 1) at the indicated time and protein level was analysed by western blot. (D) Both WT and KO RD cells were infected with ZIKV (MOI = 1) for the indicated time. Cells were fixed and stained with anti-NS1 (green) fluorescence antibody and DAPI (blue) and visualized under confocal microscope. (E) Both wild type RD cells and ERp57-KO cell line were transfected with NS1-flag plasmid or vector, then infected with ZIKV (MOI = 1) for 48 h. Protein samples were blotted to investigate the expression level of NS1 and other DDR related factors. (F) Western blot images of WT and ERp57-KO cells transfected with NS1-Flag plasmid or vector for 24 h.

Article Snippet: Anti–βactin, anti-GAPDH, anti-ERp57 and other antibodies for apoptosis factors were purchase from Santa Cruz, anti-H2AX, anti-ATM and anti-p-ATM antibodies were purchased from Cell Signaling Technology, rabbit anti-ZIKV NS1 and other nonstructural protein antibodies were purchased from GeneTex, anti-Flag tag antibodies (Sigma-Aldrich) were also used for detection at the appropriate dilutions.

Techniques: Expressing, Staining, Infection, Virus, Fluorescence, Microscopy, Knock-Out, Western Blot, Transfection, Plasmid Preparation