FIG 8.

HCMV UL138 colocalizes and interacts with STING during HCMV productive infection with a clinical strain virus. (A) NHDFs mock infected or infected with the indicated TB40/E-GFP virus at an MOI of 1 for 48 h were stained for FLAG-tagged UL138 and endogenous STING. Nuclei were counterstained with Hoechst. GFP served as a marker for viral infection (n = 3). (B) Coimmunoprecipitation experiments with NHDFs mock infected or infected with the indicated virus at an MOI of 3 for 24 h, with Western blotting performed for the indicated proteins (n = 3). (C) NHDFs mock infected for 24 h or infected with the indicated virus at an MOI of 1 and harvested at the indicated hour postinfection, with Western blotting performed for the indicated proteins. Tubulin served as a loading control (n = 3). (D) Quantitation of STING protein levels from panel C normalized to tubulin levels and shown relative to mock-infected controls from the same blot (n = 3). (E) NHDFs mock infected or infected with the indicated virus at an MOI of 1 for 24 h were analyzed for STING transcripts by RT-qPCR. STING transcript levels were normalized to GAPDH transcripts and are shown relative to mock-infected cells from the same experiment (n = 3). (F) NHDFs mock infected for 24 h or infected with the indicated virus at an MOI of 1 and harvested at the indicated hour postinfection, with Western blotting performed for the indicated proteins. Tubulin served as a loading control (n = 3). (G) Quantitation of STING protein levels from lysates used for panel F normalized to tubulin levels and shown relative to mock-infected controls from the same blot (n = 3). (H) NHDFs mock infected or infected with the indicated virus at an MOI of 1 for 24 h were analyzed for STING transcripts by RT-qPCR. STING transcript levels were normalized to GAPDH transcripts and are shown relative to mock-infected cells from the same experiment (n = 3). Bar graphs show the means ± SEM from the indicated number of biological replicates.