Figure 5
From: Human Polycomb groupEED protein negatively affects HIV-1 assembly and release
RNA interference targeting endogenous EED. (a), Experimental protocol. 293T cells were transfected with a constant amount (3 μg/106cells) of a mixture of pSUPER + pSUPER-i-EED at various ratios of each plasmid, and posttransfected with pNL4-3Luc(R-E-) 24 h later. Virus yields were determined in culture medium after a further 48 h incubation. (b) Immunofluorescence (IF) signal of endogenous EED proteins in cells transfected with control pSUPER (upper panel) or pSUPER-i-EED (lower panel) at 3 μg/106cells each, and reacted with anti-EED antibody (1:200) and FITC-labeled conjugate(1:320). (c, d), Virion production was monitored by genomic RNA levels (c), and CAp24 immunoassays (d). Virus production increased in parallel with the decrease of EED signal and as a function of pSUPER-i-EED input, with a maximum of 4- to 5-fold over the control.