Figure 4.

Investigation of the interaction between GRP78 and ZIKV E protein. A549 cells were mock-infected or infected with ZIKV-T or ZIKV-U at MOI 5 or 2, respectively and cell lysates were collected on day 1 post infection. (A) Immunoprecipitation was performed using a pan specific anti-flavivirus E protein antibody (HB112) for pull down followed by western blot analysis with an anti-GRP78 antibody. To confirm the immunoprecipitation of ZIKV E protein, the membrane was re-probed with anti-ZIKV E antibody. (B) Reverse co-immunoprecipitation was performed using an anti-GRP78 antibody for pull down followed by western blot analysis with an anti-flavivirus E protein antibody. The membrane then re-probed with an anti-GRP78 antibody to confirm the immunoprecipitation of GRP78 protein. For (A) and (B) repeat probings are separated by a continuous black line and uncropped blots are presented in Supplemental materials.