Anti mouse igg fluorescein isothiocyanate fitc

Cells were seeded on cover slips at least 2 days prior to irradiation and were fixed in 70% EtOH for 10 min at the indicated time points after exposure. Cells were permeabilised in 0.2% Triton-X in phosphate-buffered saline (PBS) and incubated with anti-phospho-Histone H2AX antibody Ser139 (Upstate/Millipore) followed by secondary anti-mouse IgG fluorescein isothiocyanate (FITC) (Sigma-Aldrich), both in PBS containing 2% bovine serum albumin (BSA). Coverslips were counterstained using DAPI and mounted on an objective glass in Vectashield. Photos were taken, and the total number of γH2AX foci per cell was scored in 50 cells per treatment and experiment using a macro for ImageJ version 1.43u, as previously described [30 (link)].

Cells that were grown on coverslips under the indicated conditions were fixed in 4% formaldehyde at 20°C for 15 min and permeabilized in methanol at −20°C for 10 min. The coverslips were then incubated at 20°C for 3 h with anti-HCV core protein polyclonal (catalog no. ab58713; Abcam) (1:200) and anti-HBx monoclonal (catalog no. sc-57760; Santa Cruz) (1:500) antibodies (Fig. 1C and D; see also Fig. S1D at https://dv.pusan.ac.kr/SynapDocViewServer/viewer/doc.html?key=402882e382e741b101845c07289e5ac8&convType=html&convLocale=ko&contextPath=/SynapDocViewServer/) and with anti-HA polyclonal (catalog no. ab9110; Abcam) (1:200) and anti-HCV core protein monoclonal (catalog no. ab-2740; Abcam) (1:200) antibodies (Fig. 2D). The cells were then incubated with anti-rabbit IgG–rhodamine (catalog no. 31670; Invitrogen) (1:200 dilution) and anti-mouse IgG–fluorescein isothiocyanate (FITC) (catalog no. F0257-1ML; Sigma-Aldrich) (1:100 dilution) antibodies at 20°C for 1 h. The slides were prepared with UltraCruz mounting medium (catalog no. sc-24941; Santa Cruz Biotechnology) and visualized using an Eclipse fluorescence microscope (Nikon).