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Fig. 1 | Virology Journal

Fig. 1

From: Histone deacetylase 1 interacts with HIV-1 Integrase and modulates viral replication

Fig. 1

Identification of HIV-1 IN-associated proteins. A biotinylated DNA fragment corresponding to a part of the U3-LTR of CasBrE MuLV was immobilized on streptavidin-coupled magnetic beads. Nuclear extracts from stably HIV-1 IN-expressing 293 T cells (293 T-INsala) were incubated with beads. a After washing with 0.1 M NaCl buffer, complexes were eluted with 0.5 M NaCl buffer and resolved on a 10% SDS-PAGE. Proteins were visualized by silver nitrate staining. IN was detected by immunoblotting with anti-IN antibodies (lower panel). Marker (lane 1); 293-INsala extract (lane 2); flow-through from beads incubated with extracts (lane 3); 0.1 M NaCl washes (lane 4–5); elution (0.5 M NaCl) (lane 6). b Eluted samples from 293 T-INsala and 293 T cells (negative control) were migrated on SDS-PAGE (4–12%) and visualized by silver staining. Signals showing difference in profile between 293 T-INsala and control 293 T fractions are numbered (indicated on the right side of the gel) and were analyzed by mass spectrometry. Detection of IN by immunoblotting is presented in the lower panel. c The presence of IN and putative interacting proteins in the purified fractions from (b) along with extracts (input) from corresponding cell lines were analyzed by Western blot using specific antibodies (anti-FEN1, anti-DDX5, anti-DDX17, anti-HDAC1)

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