FIG 7.
ZAP overexpression inhibits wild-type HIV-1 and HIV-1 containing introduced CpG dinucleotides. (A and B) HeLa cells were transfected with pHIV-1, pHIV-1env86–561CpG, pHIV-1pol795–1386CpG, pHIV-1-IRES-GFP, or pHIV-1-IRES-Renilla plus either pHA-Renilla, pHA-ZAP-S, or pHA-ZAP-L. Infectious-virus production was measured using TZM-bl reporter cells infected with cell culture supernatants. Gag expression in the media, as well as Gag and Hsp90 expression in the cell lysates, was detected using immunoblotting. The bar charts show the average values of the results of four independent experiments normalized to wild-type HIV-1 plus HA-Renilla. The numbers above the bars represent the fold decrease in relative infectious-virus production due to ZAP-S or ZAP-L overexpression. (C) Bar chart and representative Western blot for ZAP expression showing the average ZAP abundance for 15 independent transfections of p-HA-ZAP-S or pHA-ZAP-L in HeLa cells. The error bars represent standard deviations. *, P < 0.05 as determined by a two-tailed unpaired t test. The black asterisks compare the virus containing introduced CpGs in the control CRISPR cells to wild-type HIV-1 in the control CRISPR cells. The red asterisks compare the virus with HA-Renilla overexpression (black bars) to HA-ZAP overexpression (red bars).