Fig. 3: Evaluation of Dolutegravir-mediated inhibition of RAG1 binding on 12RSS.
From: Biochemical activity of RAGs is impeded by Dolutegravir, an HIV integrase inhibitor
a Sequence of 12RSS containing DNA substrate used for the study. b SDS-PAGE profile for purified 6X His-tagged RAG1 nonamer binding domain (~11 kDa), marked with an arrowhead. c, d Effect of Dolutegravir on binding of RAG1-nonamer binding domain to 12RSS substrate was also assessed by incubating increasing concentration of the inhibitor (0.1, 0.3 and 0.5 mM). Products were visualised on a native PAGE and signals were detected (c). Bar graph represents quantification based on three independent repeats showing impact of Dolutegravir on NBD binding to RSS (d). Numbers for each bar corresponds to the lane numbers in the panel. (P values * 0.01 ** 0.001). e SDS-PAGE profile for purified RAG1 central domain. The central domain along with MBP tag is ~68 kDa. Protein is seen below 75 kDa marker and is marked with an arrowhead. f, g Increasing concentrations of Dolutegravir (0.1, 0.3 and 0.5 mM) was incubated with RAG1-central domain, prior to its incubation with 12RSS. Equivalent DMSO concentration was used as vehicle control in the experiment (f). Bar graph representing quantification based on three independent repeats for the same is also shown (n = 3). Numbers for each bar corresponds to the lane numbers in the panel g. (****P values <0.0001).
