Skip to main content
. Author manuscript; available in PMC: 2021 Jan 15.
Published in final edited form as: Virology. 2019 Nov 15;540:119–131. doi: 10.1016/j.virol.2019.11.010

Figure 1. Overexpression of Human GADD34 inhibits Tat-mediated HIV1-LTR activity and HIV-1 replication.

Figure 1.

MT-2 cells were mock infected or infected by spinoculation with 20 ng of p24-CA- antigen equivalent HIV-1-supernatant for indicated days. Cells were harvested for Western blot analysis using antibodies to GADD34, and CA-p24 (A) or RNA isolation and RT-PCR with GADD34 and GAPDH specific primers (B). C) HeLa CD4+ cells were transfected with 100 ng of indicated plasmid along with 100 ng of pBlue3'LTR-luc-A plasmid and 38 h later were harvested for luciferase measurement. D and E) 293FT cells were transfected with 100 ng of pBlue3'LTR-luc-A plasmid in the presence or absence of 50 ng of pcDNA3.1-TAT-1-101-FLAG (D) or 200 ng of IFIT1-Promoter-Luc plasmid (E) in the presence of 0.5 μg of indicated plasmids. Cells were harvested 38 h later for luciferase measurement. F) 293FT cells were transfected with 100 ng of pNL4-3.Luc.R-E- plasmid in the presence of 200 ng of indicated plasmids.. Cells were harvested 38 h later for luciferase measurement. G) 293FT cells were transfected with 0.25 μg of pNL4-3 plasmid in the presence of 200 ng of GADD34/Stop plasmid or indicated concentrations of GADD34 plasmid. Cells were harvested 38 h later for Western blot analysis using antibodies to HIV-1 CA-p24. H) HeLa CD4+ cells were transfected with 0.5 μg of indicated plasmids. Cells were infected 24 h later with 20 ng of CA-p24 antigen equivalent pNL.4-3/HIV-1-supernatant for 2h. The virus was removed, cells were washed with medium and infection continued for an additional 40 h before CA-p24 quantitation. I) MT-2 cells were electroporated with 2.5 μg of pNL4-3.Luc.R-E- plasmid in the presence of 5.0 μg indicated plasmids and harvested 36 h later for luciferase measurement and Western blot analysis with GADD34 specific antibodies.