Figure 5
Co-localization and interaction of PHB and DENV E protein. (A) HEK293T/17 cells were grown on cover slips and then either mock infected or infected with DENV 2 or DENV 4 at MOI 5. At 72 h post infection, the cells were fixed, permeabilized and incubated with a polyclonal anti-PHB1 antibody (RED) and pan specific anti-dengue E protein (GREEN) follow by appropriate fluorescent conjugated secondary antibodies followed by staining with DAPI (BLUE). Cells were examined under a ZEISS LSM 800 confocal microscope co-localization between PHB1 and DENV E protein shows in yellow. (B) The degree of colocalization between PHB1 and DENV E protein was assessed by determining the Pearson’s correlation coefficient. (C) The interaction between DENV E protein and PHB1 was investigated by immunoprecipitation. HEK293T/17 cells were mock infected or infected with DENV 2 or DENV 4 and at 72 h post infection protein lysates were prepared. DENV E protein was pulled down with an anti-dengue E protein antibody, and the presence of co-immunoprecipitated PHB1 determined by western blot analysis. Full, uncropped western blots can be found in the supplemental materials.
