FIG 6.

HCMV UL138 colocalizes and interacts with STING during HCMV productive infection with a laboratory strain virus. (A) NHDFs mock infected or infected with wild-type AD169 (AD) or AD169-UL138-HA (AD138HA) virus at an MOI of 1 for 48 h were stained for HA-tagged UL138 and endogenous STING. Nuclei were counterstained with Hoechst (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 (M) or infected with the indicated virus at an MOI of 1 and harvested at the indicated hour postinfection (hpi), with Western blotting performed for the indicated proteins. GAPDH served as a loading control (n = 5). (D) Quantitation of STING protein levels from panel C normalized to GAPDH levels and shown relative to the mock-infected control from the same blot (n = 5). (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). Bar graphs show the means ± SEM from the indicated number of biological replicates.