Figure 3.

Effect of INLAIs on IN catalytic activities. (A-B) IN strand transfer inhibition in ELISA assay: (A) The IN strand transfer inhibition of compounds listed in Table 1 is compared to inhibition with Raltegravir (RAL). Data represent the means of three independent experiments with standard deviations shown as error bars. (B) Additive effect of Mut101 and Raltegravir on IN strand transfer inhibition. Comparison of dose–response curves of Raltegravir alone and Raltegravir in the presence of 10 μM Mut101. Mean of triplicate with standard deviation. Dotted lines highlight the IC₅₀ of Raltegravir in both conditions (difference not significant, Student’s t-test p = 0.48). (C-D) IN 3′ processing inhibition by Mut101 and BI-D assayed using standard radioactive assay: increasing concentration of Dolutegravir (DTG, from 3.3 to 100 nM), BI-D or Mut101 (from 0.01 to 100 μM) were used. The relative cleavage efficiency is reported for BI-D and Mut101 (D), and corresponds to the ratio between the product (19 bp) and the substrate (21 bp) converted to % inhibition. DTG resulted in 16% inhibition at 100 nM. (E-F) IN Strand transfer inhibition activity of Mut101 and BI-D assayed using standard radioactive assay: increasing concentration of DTG (from 0.3 to 10 nM), BI-D or Mut101 (from 0.01 to 100 μM) were used. The relative strand transfer efficiency is reported for BI-D and Mut101 (F), and corresponds to the ratio between the strand transfer products depicted on the autoradiography and the substrate (19 bp), converted to % inhibition. DTG has an IC₅₀ of 2.7 nM.