Tritc

Cell monolayers were washed with PBS and fixed in methanol overnight at −20°C. Cell were rehydrated in PBS (3× 5min) prior to blocking in 5% Goat serum/PBST at room temperature. Cell were incubated for 1h at room temperature with specific primary antibodies targeting epitope tags; HA (Novus Biologicals); FLAG (Novus Biological); and myc-c (Novus Biologicals) antibodies at a concentration of 100 ng/ml as was antibody targeting Pan-keratin (C11) (Cell Signaling). Use of rabbit anti-GPCMV IE2 and mouse anti-GP133 was as previously described in (67 (link)). Cells were washed and incubated for 1h with secondary antibodies, either goat anti-mouse or goat anti-rabbit, congregated with FITC or TRITC (Santa cruz). Cells were counter stained with DAPI prior to mounting in anti-fade mounting medium (Vectashield).