Fig. 1

TGEV co-localizes with TfR1 and decreases its expression. A IPEC-J2 cells were infected with Dylight 488-TGEV (green), and cultured for 1 h. The cells were then stained for confocal microscopy using a rabbit anti-TfR1 pAb, followed by Dylight 633-conjugated goat anti-rabbit IgG (red). The panel shows a three-dimensional rendering of a representative field obtained using Imaris 7.2, and the arrows indicate co-localized signals (scale bar = 10 μm). b IPEC-J2 cells were infected with TGEV (MOI 5) and fixed at 1 h p.i. Cells were then stained for confocal microscopy using a rabbit anti-TfR1 Ab, followed by Dylight 633-conjugated goat anti-rabbit IgG (red) and FITC-conjugated anti-TGEV polyclonal antiserum (green). The arrows indicate co-localized signals (scale bar = 10 μm). c Indirect immunofluorescence images of TfR1 in mock- and TGEV-infected cells at 1 h p.i. and 24 h p.i. (MOI 5). Fixed cells were stained for TfR1 (red) and the nucleus was stained with DAPI (blue). Scale bar = 10 μm. d IPEC-J2 cells were uninfected or infected with TGEV (MOI 5) and harvested at different time intervals (12–72 h p.i.). The cell lysates were analyzed by western blotting using anti-TfR1, anti-TGEV-N, and anti-GAPDH antibodies as probes