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. 2017 May 10;9(5):105. doi: 10.3390/v9050105

Figure 4.

Figure 4

AME directly blocks HIV-1 integrase nuclear entry. (A) HEK293T cells were transfected with IN-EGFP. Cells were lysed, and the nucleus and cytoplasm were isolated 40 h post-transfection. IN expression, β-actin, and MCM-2 in nucleus (N) and cytoplasm (C) were measured by Western blotting (left panel); (B) The gray value of each band in (A) was quantified and analyzed by ImageJ (right panel); (C) HEK293T cells were transfected with IN-EGFP in the presence of DMSO, ivermectin, or AME. Cells were fixed, and the nuclei were stained with 4′6′-diamidino-2-phenylindole (DAPI) after 24 h. Cells were analyzed by confocal microscopy (with a 60× objective lens); (D) Ratios of fluorescence in nucleus and whole cells at 488 nm were measured from more than 25 cells per condition randomly selected from three independent experiments. The fluorescence density at 488 nm was quantified by ImageJ and statistical analysis was performed in Graphpad Prism 5.0 software. The data represent the percentage of IN-EGFP in the nucleus. Error bars represent standard deviation (SD) of all cells in each group. ** p < 0.01 (determined by Student’s t test).