Fig 8. US28 limits HCMV-induced EGFR activation during viral entry.

(A to C) Monocytes were mock-infected or infected with WT or US28Δ HCMV (MOI = 1) for 30 min. Total EGFR and phosphorylated EGFR were detected by Western blot (A) and quantified (B and C). (D) Monocytes were mock-infected or infected with WT or US28Δ HCMV (MOI = 1) for 30 min. After 30 min of uninterrupted binding and entry, monocytes were treated with AG1478 (5 μM), LY294002 (25 μM), or MK2206 (10 μM) for an additional 30 min. Phosphorylation of Akt at Ser⁴⁷³ and Thr³⁰⁸ and total Akt were detected by Western blot. (E) Monocytes were mock-infected or infected with WT, US28Δ, ΔN, or R129A HCMV (MOI = 1) for 30 min. Phosphorylated EGFR, total Akt, total Akt, phosphorylation of Akt at Ser⁴⁷³, and phosphorylation of Akt at Thr³⁰⁸ were detected by Western blot. (F and G) pSILK empty vector or pSLIK-US28-3xF-expressing THP-1 cells were treated with DOX for 24 hours and then with EGF (200 ng/mL) for 30 min. Total EGFR and phosphorylated EGFR, and FLAG (to detect US28) were detected by Western blot (F) and quantified (G). β-actin was used as a loading control. Western blots and densitometry are representative of at least 3 biological replicates per group. ns= not significant, *P<0.05, **P < 0.005 by one-way ANOVA with Tukey’s HSD post hoc test.