Fig 3. Particle-associated US28 inhibits the induction of IE1.

(A to C) Monocytes or fibroblasts were mocked-infected or infected at MOI 1 (A, B) or MOI 5 (monocytes) (C) with US28-3xFLAG HCMV for the indicated times. US28 protein and mRNA abundance were detected by Western blot (A, C) or RT-qPCR analysis (B), respectively. RT-qPCR data are means ± SEM from 4 biological replicates per group. SV40-driven mCherry expression in infected cells was assessed by Western blot for mCherry (A, C). US28-3xFLAG and US28Δ cell-free virus lysates were used as positive and negative controls for US28, respectively. The red pixels represent the over saturation of the positive control band from long exposure of the blot during image capture (C). (D, E) Monocytes were mock-infected or infected (MOI = 1) with WT, US28Δ, or US28-complemented US28Δ (US28comp) HCMV for 24 h. IE1 induction was detected by Western blot (D) and quantified (E). β-actin was used as a loading control. Western blots and densitometry are representative of at least 3 biological replicates per group. *P<0.05, ****P < 0.0001 by one-way ANOVA with Tukey’s HSD post hoc test.