FIG 6.

The introduced disulfide bonds in the HeV F TMD mutants are poorly accessible to reducing agent. (A) The samples were prepared as described in the legend of Fig. 3B, except the overlay medium consisted of BSRs in DMEM plus 10% FBS ± 6 mM TCEP. Results were normalized to levels of the samples transfected with WT HeV F and G (untreated). All data are presented as the means ± standard deviations for three independent experiments. Statistical analysis was performed using two-way analysis of variance with a Bonferroni correction. Asterisks indicate statistical significance compared to the level of the WT HeV F+G (untreated) (***, P < 0.005). (B) The samples were prepared as described in the legend of Fig. 2A, except that the samples were treated with DMEM plus 10% FBS ± 6 mM TCEP for 3 h following the metabolic label. Blue asterisks indicate trimer (F₁), purple asterisk indicates dimer (F₁), and orange asterisks indicate monomer (F₁).