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. 2019 Feb 4;13:44. doi: 10.3389/fnins.2019.00044

FIGURE 3.

FIGURE 3

HIV-1 Tat alters FOXO3 intracellular localization in SH-SY5Y cells. SH-SY5Y cells were transfected with pcDNA3.1-flag (4 μg) or pcDNA3.1-Tat-flag (4 μg) for 48 h. (A) Expression of HIV-1 Tat was measured with Western blotting. Statistical analysis is below (∗∗∗P < 0.001) (B) Statistical analysis of the mRNA level of FOXO3 as measured by qRT-PCR. (C) The expression of phosphorylated FOXO3 was analyzed with Western blotting. Statistical analysis is below (∗∗P < 0.01). (D–F) The expression of nuclear FOXO3, cytoplasmic FOXO3 and total FOXO3 was analyzed by Western blotting. Statistical analysis is below (∗P < 0.05). (G) The nuclear translocation of FOXO3 was observed by confocal microscopy using immunofluorescence staining. Red indicates FOXO3 and blue indicates nuclei. Pictures were magnified 400×. Statistical analyses of all Western blots were of at least three independent repeated experiments. Vehicle: pcDNA3.1-flag; Tat: pcDNA3.1-Tat-flag.