Fig. 5

UL38 interacts with STING (A) PK15 cells were co-transfected with empty vector (500 ng) or Myc-UL38 (500 ng) plasmids and HA-STING (500 ng) plasmids for 30 h. The cells were then lysed and immunoprecipitated with an anti-HA antibody. The whole-cell lysates (input) and immunoprecipitation (IP) complexes were analyzed by western blotting using an anti-HA, anti-Myc or anti-GAPDH antibody. (B) PK15 cells were co-transfected with empty vector (500 ng) or HA-STING (500 ng) plasmids and Myc-UL38 (500 ng) plasmids for 30 h. The cells were then lysed and immunoprecipitated with an anti-Myc antibody. The whole-cell lysates (input) and immunoprecipitation (IP) complexes were analyzed by western blotting using an anti-Myc, anti-HA or anti-GAPDH antibody. (C) PK15 cells were transfected with empty vector (500 ng) or Myc-UL38 (500 ng). At 24 h, cells were stained with anti-Myc (red) and anti-STING (green) subjected to analysis by confocal microscopy. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. (D) PK15 cells were co-transfected with empty vector (500 ng) or FLAG-UL38 (500 ng) plasmids and HA-STING (500 ng) along with Myc-STING (500 ng) plasmids for 30 h. The cells were then lysed and immunoprecipitated with an anti-HA antibody. The whole-cell lysates (input) and immunoprecipitation (IP) complexes were analyzed by western blotting using an anti-HA, anti-Myc, anti-FLAG or anti-β-actin antibody. (E) Schematic of the truncated STING. STING is truncated into two segments on average and labeled as STING-C and STING-N, respectively. (F) PK15 cells were co-transfected with Myc-UL38 (500 ng) and pCMV-HA empty vector (500 ng) or Myc-UL38 (500 ng) and HA-STING C (500 ng) plasmids or HA-STING N (500 ng) plasmid for 30 h. The cells were then lysed and immunoprecipitated with an anti-Myc antibody. The whole-cell lysates (input) and immunoprecipitation (IP) complexes were analyzed by western blotting using an anti-Myc, anti-HA or anti-β-actin antibody