Fig. 4: The ZIKV capsid protein interacts with DGAT1.
a Co-immunoprecipitation of FLAGmDGAT1 and ZIKV MR766 CStrep (-anchor). HEK293T cells were transiently co-transfected with FLAGmDGAT1 and ZIKV MR766 CStrep-expressing plasmids. Cells were lysed 2 days post transfection and clarified lysates were subjected to FLAG-specific immunoprecipitation. Lysates (Input) and immunoprecipitated (IP) samples were analyzed by immunoblotting. Note that input samples were not heated to prevent aggregation of DGAT1. Arrows indicate DGAT1-specific bands. Actin served as loading control. Shown is one representative immunoblot (n = 2). b DGAT1 co-localizes with ZIKV C at LD-surrounding membranes. Huh7 cells were transfected with a ZIKV PRVABC59 CStrep ( + anchor)-expression plasmid, fed with oleic acid the following day, and fixed after 2 days. C and DGAT1 were stained using DGAT1- and Strep II-specific antibodies. Empty vector-transfected cells served as control. LDs were visualized using BODIPY 655/676. Shown are representative images (n = 2; scale bar 10 µm, scale bar inset 1 µm).
