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Fig. 1 | Virology Journal

Fig. 1

From: Proteomic analysis of HIV-1 Gag interacting partners using proximity-dependent biotinylation

Fig. 1

Expression and localization of recombinant HIV-1 Gag. a Diagram of the expression cassette encoding MYC tagged BirA* alone and MYC-BirA* fused to the N-terminus of HIV-1. b Jurkat or HeLa cells were transfected with BirA* or BirA*-Gag for 24 h. The expression levels of BirA* and BirA*-Gag were evaluated by Western blot using anti-MYC (US Biological Life Sciences, M9601-30), anti-p24 (NIH, 183-H12-5C) and anti-actin (Abcam, ab8226) antibodies. c Immunofluorescence of Jurkat (left panels) or HeLa (right panels) cells transfected with BirA*, BirA*-Gag or the HIV-1-expressing plasmid (pNL4-3). Paraformaldehyde-fixed cells were incubated with primary anti-MYC and anti-p24 antibodies for 1 h at 37 °C and secondary antibodies Alexa Fluor 488 anti-mouse IgG and Alexa Fluor 594 anti-rabbit IgG (Molecular Probes). Coverslips were mounted in ProLong Gold antifade reagent (Molecular Probes) with DAPI to stain the nuclei. Images were acquired using a Wave FX Spinning disc confocal microscope with a 63x objective. Scale bar indicates 10 μm

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