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Figure 2 | Retrovirology

Figure 2

From: The histone chaperone protein Nucleosome Assembly Protein-1 (hNAP-1) binds HIV-1 Tat and promotes viral transcription

Figure 2

Co-immunoprecipitation of Tat with transfected and endogenous hNAP-1. A. Schematic representation of hNAP-1 structure. The acidic domains of the protein are shown by black boxes, with the indication of their boundary amino acids. The localization of nuclear export and nuclear localization signals (NES and NLS respectively) are indicated. B. Schematic representation of the regions of amino acid homology between hNAP-1 and hSET/TAF-I. C. Co-immunoprecipitation of transfected hNAP-1 with Tat. The plasmids indicated on top of the figure were transfected into HEK 293T cells. The upper two panels show western blots with the indicated antibodies after immunoprecipitation using an anti-Flag antibody; the lower two panels show western blotting controls from whole cell lysates (WCL) from transfected cells to show the levels of expression of the transfected proteins. D. Co-immunoprecipitation of endogenous hNAP-1 with Tat. The experiment was performed by transfecting HEK 293T cells with plasmids encoding GFP-Tat or GFP alone, followed by co-immunoprecipitation with anti-GFP antibody. GFP-Tat retains full transcriptional and trafficking capacities as wt Tat [69, 74, 75]. E. GST-pulldown experiment using GST-Tat and HEK 293T whole cell lysates. GST-Tat, but not control GST protein, pulled down endogenous hNAP-1.

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