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Figure 2 | Retrovirology

Figure 2

From: Virus-producing cells determine the host protein profiles of HIV-1 virion cores

Figure 2

“Spin-thru” purification allows isolation of viral cores for proteomic analysis. A, B – “Spin-thru” purification separates viral cores from extracellular vesicles. Western blot analysis with anti-CD45 mouse monoclonal antibody of the virus samples before (left two lanes) and after (right two lanes) “spin-thru” purification. CD45 was used as an extracellular vesicle marker, p24Gag as a reference viral protein (A). Western blot of culture media from uninfected 293 T/17 cells and suspension of virus harvested from pNL4-3 transfected 293 T/17 cells before and after “spin-thru” centrifugation with anti-RHA rabbit polyclonal antibody (B). C – “Spin-thru” purification separates viral cores from the envelope: Western blot analysis of VSV-G glycoprotein in the samples of VSV-G-pseudotyped HIV-1 before and after “spin-thru” centrifugation. D – Representative SDS-PAGE profiles of the preparations of purified virion cores. Coomassie blue-stained preparative SDS-PAGE 12.5% gels of HIV-1 viral cores and the cell culture supernatants of non-infected cells after “spin-thru” centrifugation were sectioned as shown by the dashed lines. Infected samples are marked as (+) and uninfected as (−). The molecular mass markers are indicated on the left and the gel fractions are specified on the right. Positions of HIV-1 core proteins p24CA and p31IN are shown on the left side.

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