CFAV inhibits DENV-1 and ZIKV replication in vitro. (A) The schematic summarizes the experimental design of the in vitro studies and shows the different treatments and controls with their inoculation time line. C6/36 cells were inoculated with DENV at 0, 2, or 3 days after CFAV infection or with ZIKV at 3 days after CFAV infection. (B) Growth curves represent CFAV RNA levels on a log10 scale measured by absolute RT-qPCR in the presence (blue filled dots) or absence (red filled dots) of DENV-1. (C and D) Infectious titers of DENV-1 (C) and ZIKV (D) over time in log10-transformed focus-forming units (FFUs) per milliliter of C6/36 cell culture supernatant in the presence (red filled dots) or absence (blue filled dots for DENV-1 or yellow filled dots for ZIKV) of CFAV. One unit was added to the raw FFU values to allow log10 transformation of zeros. Day 0 represents the day of arbovirus inoculation, where arbovirus titers are set to 0 for visualization purposes. Arbovirus titration was performed in C6/36 cells. Each data point represents the mean for 3 (CFAV and DENV-1) or 2 (ZIKV) biological replicates, and the vertical bars are the standard errors of the means. The P values above the graphs were obtained from the full statistical models. Asterisks show the statistically significant differences between each treatment at each time point and the single-virus inoculation control at the same time point (*, P < 0.05; **, P < 0.01; ***, P < 0.001).