Figure 3 GA inhibits ZIKV infection. (A) Viability of NHA infected with ZIKV and treated with GA. NHA were grown in 24-well plates to >80% confluency and treated with GA (0–20 µM) or DMSO for 3 hours and infected with ZIKV strain PRVABC59 at an MOI of 0.3. At 12 hpi, supernatants were carefully removed and replaced with fresh AGM. (B) 7 days post infection, samples were analyzed for cell viability by the MTS assay and then live cells were harvested for RNA and processed with Taqman based real-time PCR to quantify ZIKV RNA. Experiments were performed in duplicate three independent times and the data were analyzed by t-test (* indicates p ≤ 0.05). (C) Cell free ZIKV in 199V medium was pretreated with 10 µM GA or with DMSO for 1 hour at 37 °C. The pretreated ZIKV was used to infect Vero cells grown in 6 well plates >80% confluency with 0.5 MOI for 1 hour. The infected cells were washed twice with warm DPBS and supplemented with DMEM supplemented with 5% FBS. Supernatant was collected at 4, 24 and 48 hours post-inoculation for cell free viral RNA copy number determination. Experiments were performed in triplicates.