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. 2018 Sep 20;46(21):11539–11552. doi: 10.1093/nar/gky851

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — The Rev/RRE axis favors the recruitment of CBP80 and eIF4AI to the HIV-1 unspliced mRNA. (A) HeLa cells were transfected with 1 μg pNL4.3-wt or 1 μg pNL4.3-CTE together with 1 μg of pCMV-myc-CBP80 or pCMV-myc-eIF4E. At 24 hpt, the interaction between the unspliced mRNA and CBP80 or eIF4E was analyzed by ISH-PLA. Scale bar 10 mm (upper panels). Dots per cell quantifications for total unspliced mRNA-CBP80 or unspliced mRNA-eIF4E interactions are presented in the lower panels. Dots quantifications were performed with ImageJ (****P < 0.0001, Mann–Whitney test). (B) HeLa cells were transfected with 1 μg pNL4.3-wt or 1 μg pNL4.3-CTE together with 1 μg of pCIneo-HA-eIF4AI. At 24 hpt, the interaction between unspliced mRNA and eIF4AI was analyzed by ISH-PLA. Scale bar 10 mm (upper panel). Dots per cell quantifications for total unspliced mRNA-eIF4AI interactions are presented in the lower panel. Dots quantifications were performed with ImageJ (***P < 0.001 and NS; non-significant, Mann–Whitney test). (C) HeLa cells were transfected with 1 μg pNL4.3-wt or 1 μg pNL4.3-CTE together with 1 μg pCMV-myc-CBP80 and 1 μg pCIneo-HA-eIF4AI. At 24 hpt, the interaction between HA-eIF4AI and myc-CBP80 was analyzed by PLA (upper panels). Scale bar 10 μm. Dots per cell quantification of eIF4AI and CBP80 interactions were performed with ImageJ (****P < 0.0001, Mann–Whitney test). (D) HeLa cells were transfected with 1 μg pCMV-myc-CBP80 and 1 μg pCIneo-HA-eIF4AI together 1 μg pEGFP-Rev (pEGFP was used as a control). At 24 hpt, the interaction between HA-eIF4AI and myc-CBP80 was analyzed by PLA (left panels). Scale bar 10 μm. Dots per cell quantification of eIF4AI and CBP80 interactions were performed with ImageJ (***P < 0.001, Mann–Whitney test).