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Figure 4 | Journal of Biology

Figure 4

From: Endophilins interact with Moloney murine leukemia virus Gag and modulate virion production

Figure 4

Copurification of HA-endophilin 2 with virions. The 293T cells were cotransfected with 2 μg of plasmid encoding HA-endophilin 2 and 10 μg of proviral DNA. Virion particles purified through a 25/45% sucrose gradient were reloaded on a 20-60% linear sucrose gradient, and 20 fractions were collected. (a) A plot of the gradient density. (b,c) Proteins in these fractions were precipitated with TCA and analyzed by western blotting with (b) anti-HA and (c) anti-capsid antibodies. The virions migrate near the middle of the gradient and are marked by the comigration of CA and a doublet of endophilin proteins (fractions 9-11). A small amount of endophilin is also found at the top of the gradient (fractions 16-19). The bulk of the membrane-associated and low molecular weight proteins remains at the top of the gradient; the huge amount of protein in these fractions cause distortions in the mobility of the proteins in these lanes. High-molecular-weight proteins at the top of the gradient are also recognized by weak nonspecific reactivity in the anti-HA antibody.

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