Goat anti mouse coralite 594 antibodies

HeLa cells or HEK293T cells grown on coverslips in 6-well plates were transfected with corresponding plasmids for 48 h. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and blocked with phosphate-buffered saline (PBS) containing 10% goat serum and 0.1% Triton X-100. The cells were then incubated with the indicated primary antibodies at 4°C overnight and with goat anti-mouse-CoraLite 594 antibodies (Proteintech, China) for 1 h. Nuclei were stained with DAPI (Bestbio, China). The IRF3 antibodies were purchased from Proteintech (China). Images were acquired with a Zeiss LSM880 confocal microscope (Germany) and analyzed with ZEN (blue version) and ImageJ.

At different time points after viral infection, RD cells were fixed with precooled methanol for 5 min and then permeabilized with 0.1% Triton X-100 for 5 min. The cells were blocked with 10% goat serum in PBST for 1 h, incubated with rabbit anti-EV-A71 VP1 and mouse anti-PHB2 antibodies overnight at 4 °C, and incubated with goat anti-rabbit-CoraLite 488 and goat anti-mouse-CoraLite 594 antibodies (Proteintech) for 1 h. Nuclei were stained with DAPI (BestBio, Shanghai, China). Images were acquired using an Olympus FluoView FV10i confocal microscope (Olympus, Tokyo, Japan) and analyzed with FV10-ASW 3.0 Viewer software (Olympus).