Figure 5

Activation of NF-κB involves PI3K/Akt signaling in HIV-1 Vpr-mediated induction of CCL5 in astrocytes. SVGA astrocytes were pre-treated with the chemical inhibitor for the PI3K/Akt pathway (LY294002) 1 h before mock transfection or transfection with a plasmid encoding HIV-1 Vpr. (A) and (B) Effect of LY294002 on mRNA expression and protein concentration for CCL5, respectively. SVGA cells were also transfected with siRNA targeting Akt isoforms (Akt-1, Akt-2 and Akt-3) followed by mock transfection or transfection with a plasmid encoding Vpr. (C) and (D) Effect of siRNAs on CCL5 mRNA expression and protein concentration, respectively. (E) Specificity of siRNA for Akt isoforms against individual subunits. (F) Inhibition of NF-κB (p50) nuclear translocation with LY294002. The bars represent the mean ± SE of three independent experiments done in triplicate. Statistical significance was determined using Student’s t-test, **p < 0.01, *p < 0.05