Extended Data Fig. 2: Transiently-expressed PRV pUL36 interacts with components of the Kif5 motor complex.

(A) pUL36 region 7 is dispensable for co-immunoprecipitation (co-IP) with Kif5 heavy chain (KHC) and Kif5 light chain (KLC). HEK293 cells were lysed 16-18 hpt and GFP-pUL36 was immunoprecipitated (IP) with anti-GFP antibody and detected by western blot as indicated. Inputs are 6% of crude lysates (n=3 independent experiments) . (B) Peripheral accumulations of transiently expressed GFP-pUL36ΔR7 co-localize with endogenous cellular Kif5 components (arrows). Vero cells were fixed and immunostained for KHC or KLC at 20 hpt (n=3 independent experiments). Scale bars are 20 µm. (C) GFP-pUL36ΔR7 co-immunoprecipitates with KLC1 and KLC2 isoforms. HEK293 cells transiently expressing GFP or GFP-pUL36ΔR7 were co-transfected with HA-KLC1 and HA-KLC2. Endogenous KHC served as a positive co-immunoprecipitation control (n=3 independent experiments). (D) Transient expression mCherry-KLC1 and mCherry-KLC2 in Vero cells. Both fusion proteins show varying degrees of juxtanuclear localization and an absence of peripheral accumulation (n=3 independent experiments). (E) mCherry-KLC1 and mCherry-KLC2 redistribute with GFP-pUL36ΔR7 to the cell periphery during transient expression. Vero cells were imaged 24-28 hpt (n=3 independent experiments). Scale bars are 20 µm.