The following antibodies were used: mouse anti-HA antibody (1:500, BioLegend), rabbit anti-Giantin antibody (1: 1000, Biolegend), rabbit anti-PDI antibody (1:50, CST), goat anti-Olig2 antibody (1:200, R&D), rabbit anti-GFAP antibody (1:200, Proteintech), rabbit anti-IBA1 antibody (1:250, Invitrogen), mouse anti-Neu-N antibody (1:100, Millipore), rabbit anti-HCoV-OC43-N antibody, Cy3-conjugated donkey anti-rabbit antibody (1:500, Jackson), Alexa Fluor 488-conjugated donkey anti-mouse antibody (1:500, Jackson), Alexa Fluor 555-conjugated donkey anti-mouse antibody (1:500, Jackson), Alexa Fluor 488 anti-HA.11 Epitope Tag Antibody (1:500, Biolegend), Cy3-conjugated donkey anti-goat antibody (1:500, Jackson).
17CL-1 Cells infected with the virus (MOI = 0.1) were fixed using 4% PFA in PBS, then the cells permeabilized with 0.2% Triton X-100 in PBS. Cells were incubated with primary antibody at RT for 1 h, and then incubated with second antibody at RT for 1 h, the nucleus was stained with DAPI. After three times wash with PBS, the coverslips were mounted on glass slides. Cells were examined by confocal microscopy (ZEISS).
17CL-1 Cells infected with the virus (MOI = 0.1) were fixed using 4% PFA in PBS, then the cells permeabilized with 0.2% Triton X-100 in PBS. Cells were incubated with primary antibody at RT for 1 h, and then incubated with second antibody at RT for 1 h, the nucleus was stained with DAPI. After three times wash with PBS, the coverslips were mounted on glass slides. Cells were examined by confocal microscopy (ZEISS).