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. 2021 Jan 19;12(1):e02525-20. doi: 10.1128/mBio.02525-20

FIG 1.

FIG 1

Establishment of HIV-dual-GT reporter cell clones. (A) Map of the HIV-dual-GT proviral reporter construct. (B) Western blot analysis of lysates of HEK293T cells transfected with HIV-dual-GT or HIV.r−e− in the presence or absence of a Rev expression plasmid (pcRev) with antibodies to the p24 capsid protein, α-tubulin, or Rev. (C) 293T cells transfected with HIV-dual-GT and pcRev or pcDNA3.1 were analyzed by flow cytometry for Gag-BFP and DsRed expression. (D) Jurkat cells transduced with the HIV-dual-GT reporter vector (MOI, 0.3) and transiently transfected with pcRev or pcDNA control plasmid were analyzed by flow cytometry. (E) Scheme for the generation of reporter cell clones. (F) HIV-dual-GT-transduced Jurkat reporter cell clones J-dual#3 and J-dual#6 were superinfected with VSV-G-pseudotyped HIV-1 viruses differing only in the expression of Rev and analyzed by flow cytometry. All assays were performed at least twice, and results of one representative experiment are shown.