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. 2009 Aug 22;1792(10):1027–1035. doi: 10.1016/j.bbadis.2009.08.009

Fig. 7.

Fig. 7

Overexpression of neither wild-type nor catalytically inactive MGRN1 significantly affects HIV-1 GAG release from cells. HEK293T cells were cotransfected with an HIV-1 GAG p55 expression construct and either a GFP control, a positive control for budding inhibition (GFP-VPS4(EQ)), or GFP fusion constructs expressing wild-type MGRN1 or catalytically inactive MGRN1 (AVVA mutant). (A) Representative immunoblots from 1 of 3 replicate experiments. Top panel: GAG IB of virus-like particle (VLP) preps from media of transfected cells. Middle panel: GAG IB of cell lysates of transfected cells. Bottom panel: non-specific bands from upper region of cell lysate blot demonstrate even loading. (B) Graphical representation of the effect of each treatment on VLP budding, averaged from 3 replicate experiments. Relative budding index (y axis) represents the ratio of GAG signal in VLPs to GAG signal in cell lysates of each transfection relative to that of GAG + GFP control samples. VPS4(EQ) reduced GAG release (p < 0.02) while neither wild-type nor catalytically inactive MGRN1 had a significant effect (p > 0.3). The standard error of the GAG + GFP transfected samples is 0 because the budding index of these samples is defined as 1 within each experiment.