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. 2021 Aug 30;9:724970. doi: 10.3389/fcell.2021.724970

FIGURE 5.

FIGURE 5

HIV Tat-mediated IRF3 phosphorylation and nuclear translocation. (A) Representative Western blot and quantification of p-IRF3, IRF3 in the cell lysates from human monocytes exposed to HIV Tat for various time points (5 min – 3 h). N = 3. (B) Representative Western blot and quantification of IRF3 in the nuclear cell lysates from human monocytes exposed to HIV Tat for various time points (5 min to 3 h). N = 4. (C) Representative Western blot and quantification of IRF3 in the cytoplasmic cell lysates from human monocytes exposed to HIV Tat for various time points (5 min to 3 h). N = 4. (D) Representative images of human monocytes transfected with IRF3-GFP plasmid, followed by HIV Tat exposure for 15 min. Scale bars: 20 μm. White arrow, IRF3 nuclear translocated cells. (E) Quantification of IRF3 nuclear translocation (Student’s t-test). N = 3. (F) Schematic illustration of IRF3 binding sequence on the promoter region of CXCR3. (G) ChIP assay demonstrating HIV Tat-mediated binding of IRF3 to the CXCR3 promoter. One-way ANOVA with post hoc test. All data are presented as mean ± SD of three or four individual experiments (Biological replicates). *p < 0.05 versus control group; **p < 0.01 versus control group; ***p < 0.001 versus control group.