Fig. 2.

Quercetin pretreatment decreases RV endocytosis and RV-induced Akt phosphorylation. BEAS-2B cells were pretreated with 10 μM quercetin or DMSO and incubated with sham, UV-RV1B, RV1B, UV-RV39 or RV39 for 30 min at 4 °C, washed to remove unbound virus and cells were further incubated for 30 min at 37 °C. Endocytosed virus was detected using antibody to RV followed by flow cytometry (A and B). Data represent average and SD calculated from three independent experiments (^∗p ⩽ 0.05, different from sham; ^†p ⩽ 0.05, different from media or DMSO treated cells). (C) Cells were pretreated with DMSO or quercetin and incubated with RV1B for 30 min. Cells were washed and incubated with antibody to RV capsid protein (green) and EEA1 antibody (red). Nuclei were counter-stained with DAPI (blue) and cells were visualized by confocal microscopy. Arrows represent co-localization of RV1B with EEA1. Cells pretreated with quercetin were infected with sham or RV1B and incubated for 30 min. Total cell lysates were subjected to Western blot analysis with antibodies to p-Akt or total Akt (D). Images and blot are representative of three independent experiments.