Evaluating Antiviral and Inhibition Effects of Compounds

To confirm the inhibition effect of Rico on drugs of abuse (Coc and Meth) and to check the antiviral efficacy of NF against HIV infectivity, we conducted the p24 antigen assay using a standard protocol. HAs (10× 10⁶ cells) were infected with HIV-1Ba-L (NIH AIDS Research and Reference Reagent Program, # 510) overnight as described earlier (V. S. Atluri, et al., 2013 (link)), washed with PBS (pH 7.4), and further incubated with and without Coc+Meth treatment, plus NF treatment at a concentration of 100 µg/mL. The culture supernatants from days 1^st, 2^nd, 3^rd, 5^th, and 8^th day after 5 DPI (total 14 of HIV infection) were quantified for p24 antigen (pg/ml) using p24 ELISA kit (ZeptoMetrix, NY).