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. 2017 Mar 22;8(24):39922–39934. doi: 10.18632/oncotarget.16485

Figure 4. HIV-1 Tat downregulates EAAT-2 in U87 cells via AEG-1 in U87 cells.

Figure 4

U87 cells were transfected with pRK5M-flag (3 μg), pRK5M-Tat-flag (3 μg), pcDNA3.1-myc (3 μg), pcDNA3.1-AEG-1-myc (3 μg), Pll3.7-AEG-1-scamble (3 μg), or Pll3.7-AEG-1-shRNA (3 μg) for 48 h. A.-B. HIV-1 Tat protein expression was increased in pRK5M-Tat-flag-transfected cells compared with pRK5M-flag-transfected cells. C.-D. AEG-1 protein expression was increased in pcDNA3.1-AEG-1-myc-transfected cells compared with pcDNA3.1-myc-transfected cells. Also, AEG-1 protein expression was decreased in Pll3.7-AEG-1-shRNA-transfected cells compared with Pll3.7-AEG-1-scamble-transfected cells. E.-H. EAAT-2 protein expression was decreased after overexpression of HIV-1 Tat or AEG-1 in U87 cells, suggesting that there is a synergistic effect on EAAT-2 regulation. The ability of HIV-1 Tat to downregulate EAAT-2 was inhibited by AEG-1 knockdown in conjunction with HIV-1 Tat overexpression. Data are plotted as mean ± SEM (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001. Null: untreated; Ctr: pRK5M-flag or pcDNA3.1-myc; Tat: pRK5M-Tat-flag; AEG-1: pcDNA3.1-AEG-1-myc; Ctr-shRNA: Pll3.7-AEG-1-scamble; AEG-1-shRNA: Pll3.7-AEG-1-shRNA.