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. 2012 Jul 12;23(9):1003–1015. doi: 10.1089/hum.2012.046

FIG. 2.

FIG. 2.

Equol and resveratrol enhance viral uptake but attenuate viral replication. (A) Cells were infected with the nonreplicating Ad5GFP mutant alone (22Rv1, 25 ppc; DU145, 50 ppc; PC-3, 300 ppc) and in combination with equol (100 μM) or resveratrol (10 μM) and analyzed by flow cytometry 48 hr later. Results are percent GFP-positive cells in combination-treated versus AdGFP infected alone; averages±SD, n=3. (B) Cells were infected with wild-type virus (Ad5) (22Rv1, 25 ppc; DU145, 50 ppc; PC3, 300 ppc) and treated with equol or resveratrol as above. Viral E2A DNA was quantified by quantitative PCR (qPCR) 4 hr after infection. Data are normalized to cellular GAPDH DNA in each sample and to viral E2A DNA in cells infected with Ad5 alone, averages±SD, n=3. (C) Equol (100 μM) and resveratrol (10 μM) increase expression of cell surface receptors essential for adenoviral entry. The proportion of receptor-positive cells was determined by flow cytometry 24 hr after treatment, averages±SD, n=3. (A–C) Data analyzed in each cell line by one-way ANOVA with Bonferroni multiple comparison test, compared to untreated cells. (D) Viral DNA amplification over time (3–72 hr) was determined by qPCR analysis with primers to hexon DNA in 22Rv1, DU145, and PC-3 cells infected with Ad5 (100 ppc) with and without the addition of equol (E, 100 μM) or resveratrol (R, 10 μM). Results are normalized to cellular actin DNA in each sample and to viral DNA present 3 hr after infection with Ad5 alone, averages±SD, n=3. (E) Viral replication over time (3–72 hr) was determined by TCID50 assays in samples treated as described above for 22Rv1, DU145, and PC-3 cells. Results are averages±SD, n=3. (D–E) Two-way ANOVA with Bonferroni post-tests comparing each result over the time period to Ad-infected untreated cells for each time point. (F) Cells infected with Ad5 (22Rv1 and DU145, 100 ppc; PC-3, 300 ppc) with and without simultaneous additions of equol (E, 100 μM) or resveratrol (R, 10 μM) analyzed for viral E1A mRNA and protein expression 24 and 48 hr later, averages±SD, n=2–3, representative immunoblots.