Figure 5
Profiling of anti-HIV activity during iterative fractionation of Cistus incanus (Ci) extract reveals multiple active compounds.
(A) Anti-HIV activity profile of 45 high-activity fractions obtained during initial separation of Ci extract. The high-activity fractions A4-D12 were identified by single-dose anti-HIV screening of a set of 96 fractions generated in the initial separation step. Screening was performed with LC5-RIC cells and HIV-1LAI. For quantification of anti-HIV activity of each high-activity fraction, cells were exposed to the virus in the presence of different concentrations of the fraction, ranging from 100 μg/ml to 3.125 μg/ml (2-fold serial dilutions) and EC50 values determined. Symbols signify mean EC50 values of triplicate assays for each fraction and error bars the standard deviation of the mean. The frame marks an exemplary activity cluster, consisting of fractions A8 –A12. (B) Anti-HIV activities of fine fractions derived from the exemplary activity cluster identified in A (fractions A8-A12). The activity cluster was subjected to a second fractionation step, yielding fine fractions A-M, which were then assayed for anti-HIV activity. Bars indicate the mean values of 3 independent assays for each fraction and error bars the standard deviation of the mean. #: No antiviral activity detected in the measured concentrations. (C) Anti HIV activities of high-resolution fractions derived from fine fraction F. Fine fraction F was subjected to a third fractionation step, yielding high-resolution fractions F1 to F9 which were again assayed for anti-HIV activity. Bars indicate the mean values of 3 independent assays for each fraction and error bars the standard deviation of the mean. #: No antiviral activity detected at the highest concentration.
