FIG 4.

Colocalization of gH mutants with the endoplasmic reticulum. RK13 cells were cotransfected with expression plasmids for wild-type (WT) or mutated gH in combination with gL and the ER marker pmTurquoise2ER (Addgene). At 24 h posttransfection, cells were fixed with 3% paraformaldehyde and permeabilized with 0.1% Triton X-100. gH was detected using a monospecific rabbit antiserum and Alexa Fluor 633-conjugated secondary antibodies. Green (ER) and red (gH) fluorescence was excited at 488 nm and 633 nm, respectively, and recorded with a laser scanning confocal microscope (Leica SP5). Channels were merged to visualize ER localization of gH. Size bars, 10 μm.