Associations with F are altered by O-glycan loss.

A) Cell lysates of cells transfected with wt F and either wt or mutant HeV G. HeV G (top) is blotted with rabbit anti-HA, and HeV F (bottom) is blotted with mouse anti-AU1. B) Pulldown of HeV G and co-IP of F for transfected cells. Antibody blotting was performed as in A). C) Cell lysates of cells transfected with wt F and wt or mutant NiV G. D) Co-IP of NiV transfected cells. E) Co-IP values of HeV and NiV O-glycan mutants, determined by densitometry. Image Lab software (Biorad) was used to measure the densitometry of Western blot bands. Co-IP values were determined by dividing either the F0, F1, or total F densitometry values for the co-IP bands by the co-IP G densitometry values to account for any differences in co-IP G pulldown. These values were then divided by F0, F1, or total F densitometry values for cell lysates to account for differences in F mutant expression. Values were then normalized to those of wt HNV G. Averages are shown. N = 3. Statistically significant differences, as determined by a Student’s t-test, are marked with an * (p<0.05).