Fig. 3.

CHIKV TF forms homo-oligomer. (A) DTT reduced the oligomerization of CHIKV TF. Constructs expressing TF-Flag or TF-C/A-Flag were transfected to 293T cells. The cells were incubated with 50 μM 2-BP for 6 h before isolation. The cell lysates were loaded with protein loading buffer containing no DTT, 10 mM DTT, or 50 mM DTT for Western blot analysis. (B) Prediction of disulfide bonds in TF. TF from CHIKV strain 181/25 was predicted by DiANNA 1.1 web server (http://clavius.bc.edu/∼clotelab/DiANNA/). (C) Diagram of TF-C/A mutant. (D) TF-C/A mutation reduced the association of TF compared with wild type TF. Constructs expressing GFP, GFP-TF, or GFP-TF-C/A combined with TF-Flag were transfected to 293T cells. GFP-Trap immunoprecipition was performed and accessed by Western blot with indicated antibodies.