Fig. 4: PU.1 levels decrease in the presence of Vpr.

A NL4-3 ∆GPE-GFP viral genome map. B Representative flow cytometry histogram of PU.1 expression in infected (GFP⁺) or uninfected bystander (GFP^-) MDMs infected with NL4-3 ∆GPE-GFP with or without vpr and collected on day 7 post infection. C Summary graph showing the percentage of infected (GFP⁺) cells that do not express PU.1 as determined by flow cytometry as depicted in Fig. 4B. The mean ±standard deviation from n = 4 independent donors is shown for each time point. P values were determined using a two-sided, one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test. ***p < 0.001; ****p < 0.0001. D Lentiviral map of vectors encoding 3xFLAG-encoding vpr or vpx genes. E PU.1 expression plasmid map for full-length, human PU.1. F Representative flow cytometric plots of HEK 293T cells transiently transfected with PU.1 and the indicated FLAG-tagged viral protein. G Summary graph showing PU.1⁺ expression in transfected (FLAG⁺) cells. Each point represents the mean of three technical replicates. The mean ± standard deviation of four independent experiments is shown. P values were determined using one-way ANOVA compared to control; ****p < 0.0001. H Immunoblot analysis of PU.1 in K562 cells transduced with the indicated lentivirus expressing 3xFLAG-tagged viral proteins. Results are representative of those from three independent experiments. I Summary graph of SPI1 (the gene encoding PU.1) expression in HEK 293T cells co-transfected with PU.1 and an 89.6 expression vector with or without an intact vpr open reading frame. SPI1 levels were assessed from purified RNA using RT quantitative real time PCR. Results represent the mean fold change compared with wild type ±standard deviation for samples performed in triplicate for two independent experiments. Source data are provided as a Source Data file.