Figure 3
Comparison of effects of Nef-WTs and Nef-CPs on enhancement of viral infectivity. pBRmac239Δnef/luc was cotransfected into HEK293 cells with either pNefmac239-WT, pNefmac239-CP, pNefmac239-G2A, pNefmac239-CP-G2A, or an empty vector (A). pBRmac239Δnef/luc was cotransfected into HEK293 cells with either pNefJR-CSF-WT, pNefJR-CSF-CP, or an empty vector (B). pNL-CHΔenv Δnef/luc and psvJR-FLenv were cotransfected into HEK293 cells with either pNefJR-CSF-WT, pNefJR-CSF-CP, or an empty vector (C). pNL-CHΔenv Δnef/luc and psvJR-FLenv were cotransfected into HEK293 cells with either pNefmac239-WT, pNefmac239-CP, or an empty vector (D). After a 72 h cultivation, the cells were collected and subjected to SDS-PAGE and western immunoblot analysis to detect Nef and β-actin, as described in Materials and Methods (top panels). The supernatants were subjected to SIV p27 or HIV-1 p24 enzyme-linked immunosorbent assay (ELISA) and infectivity assay using MAGIC-5 cells, as described in Materials and Methods (bottom panels). Each bar represents the mean standard deviation (n = 3).