In Vitro Inhibition of HIV1-D1-A2 pre-mRNA Splicing by IDC16 and Complementation with SR Proteins SC35 and SF2/ASF

<div><p>(A) Schematic representation of HIV1-D1-A2 transcript is shown. Numbering of the HIV-1/BRU RNA sequences is according to Ratner et al. [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0030159#ppat-0030159-b049" target="_blank">49</a>].</p><p>(B) Polyacrylamide gel electrophoresis of the in vitro splicing products of HIV1-D1-A2 transcript. Uniformly labeled RNA was prepared as described under Materials and Methods and was incubated alone (lane 1) in a HeLa cell nuclear extract under splicing conditions in the absence (lane 2), in the presence of 10% DMSO (lane 3), or 5 μM, 10 μM, 20 μM, and 40 μM of compound IDC16 (lanes 4–7, respectively).</p><p>(C) The same reactions as in (B) lanes 3–7 were complemented with 0.21 μM of recombinant SF2/ASF (lanes 1–5, respectively).</p><p>(D) The same reactions as in (B) lanes 3–7 were complemented with 0.19 μM of recombinant SC35 (lanes 1–5, respectively).</p><p>(E) Splicing products of the Minx transcript analyzed by polyacrylamide gel electrophoresis. Uniformly labeled RNA was incubated alone (lane 1), in HeLa nuclear extract in the absence (lane 2) or presence of 5 μM, 10 μM, 20 μM, 30 μM, or 50 μM of compound IDC16 (lanes 3–7), respectively.</p>