Figure 2.

Co-immunoprecipitation and pull-down of recombinant capsid and DDX3X. (A) Full length His-capsid and α2-4 His-capsid were transfected in HEK293T cells and lysates were prepared 24 h p.t. Lysates were used for IP with capsid antibody and blots were probed for capsid, DDX3X and GAPDH. (B) Map of the HA-DDX3X deletion constructs. (C) Total lysates were prepared from HEK293T cells transfected with HA-DDX3X-FL and HA-DDX3X deletion constructs at 24 h p.t. Purified His-capsid was added to the lysate for in-solution interaction and protein complexes were pulled down by Ni-NTA beads. Lysates incubated with Ni-NTA beads only were used as negative control. The beads were washed, boiled in Laemmli buffer and loaded on SDS PAGE and probed for HA-tag, capsid and Actin. In the first lane of blot (i) purified His-capsid was loaded as a positive control for capsid antibody. Total lysate of HEK293T cells transfected with HA-DDX3X-Δ351-661 and total lysate of HEK293T cells transfected with HA-DDX3X-Δ351-661 and infected with DENV was loaded as positive controls respectively in blots (ii) and (iii) for HA, capsid and actin antibodies.