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. 2020 Jun 12;119(2):419–433. doi: 10.1016/j.bpj.2020.05.035

Figure 2.

Figure 2

Membrane relocalization of HIV-1-gRNA by Gag. (A) HeLa cells stably expressing MS2-eGFP were transfected with a construct encoding pIntro (see Materials and Methods). Nuclei were stained with Hoechst33258 (blue channel, column 1). Nontransfected cells showed GFP signal in the nuclei and nucleoli, whereas in cells transfected with pIntro, the MS2-eGFP fluorescence signal was only localized in the nuclei but no more in the nucleoli (green channel, column 2). The merge is in column 3. (B) The cells were then transfected with a plasmid encoding for Rev. This cotransfection ensured the complete export of the MS2-eGFP-labeled gRNA from the nucleus to the cytoplasm because of the specific recognition of the RRE. When Gag alone (C) or in mixture with Gag-mCH (D) was coexpressed, gRNA was relocalized to the PM. Confocal microscopy was performed 24 h post-transfection. Cartoons on the right illustrate the observed localizations of MS2-eGFP-RNA. Cyt, cytoplasm; N, nucleus; NU, nucleolus. To see this figure in color, go online.