Fig. 1

A HSV-1 infectious viral particles (ivp) released from T₃ treated latently infected mouse TG explants. TGs from mice n = 10 latently infected with HSV-1 were explanted 30 days post infection. TG explants were separated into replicates of two treatment groups, +T₃ and −T₃, and were cultured for 8 days post explant. Media from each replicate was quantitatively tested daily for HSV-1 ivp via plaque assay. Two-way ANOVA with Holm-Sidak post hoc analysis suggests that statistically significant differences in ivp between +T₃ and −T₃ treatment at days 6, 7, and 8. Asterisk denotes p < 0.001. B PI3K Pathway is active in differentiated LNCaP cells with pAkt increase in differentiated cells. Western blot was performed using rabbit monoclonal IgG antibody against phospho-AKT pSer473 (ThermoSci, Cat#: OMA-03061) and mouse antibody AKT (Rockland, Cat#: 200-301-401) at a dilution of 1:1000 followed by the addition of conjugated secondary antibody for detection on extract from undifferentiated and differentiated LNCaP cells. C PI3K inhibitor reversed T₃-mediated repression HSV-1 viral replication from differentiated LNCaP cells treated with 100 nM T₃ and/or 20 µM LY294002 (Sigma Aldrich, cat#: L9908) was measured quantitatively by FLICIT assays [68]. In short, Vero cells were seeded on 384-well plates followed by exposure to media from EGFP HSV-1 infected cultures. The infected media samples were applied in serial dilutions in replicates and were incubated for 8–18 h when EGFP was observed. The numbers of total cells and infected cells were imaged and quantified by the BioTek Cytation3 fluorescent imaging station and Gen5 software then used to calculate the viral titer using an inverse Poisson’s equation as previously described. Two-way ANOVA with Holm-Sidak post hoc analysis suggests that statistically significant differences in fluorescently labeled infected cells per mL exists. a p < 0.018, b p < 0.004, c p < 0.012, d p < 0.035