gp120-mediated increase of IL-6 expression in SVGA astrocyte cells.

<p>1×10⁶ SVGA astrocytes were transfected with 1 µg gp120 JR-FL (R5 tropic) DNA using Lipofectamine™ 2000. The cells were harvested at the times described in the text. Mock transfection was performed with transfection of equal amount of empty human vector pcDNA3.1. All times noted on the figure and listed in the text are the times at which cells and supernatants were harvested after the end of the 5 hour transfection protocol. IL-6 mRNA (A) and IL-6 protein (B) expression levels were measured using real time RT-PCR and bioplex assays respectively. In (B), open bars show empty-vector transfected mock controls and closed bars show gp120 transfected samples. (C) shows the effect of exogenous gp120 on SVGA cells. SVGA were exposed to 20 nM recombinant gp120IIIB for various lengths of time and total mRNA was isolated. IL-6 expression levels were measured using real time RT-PCR. The mRNA levels are presented as fold difference between gp120 transfected cells and control cells transfected with empty plasmid. The protein concentration is presented as ng/ml protein in supernatant. Each bar represents mean ± SE of 3 experiments with each experiment done in triplicates. The statistical significance was calculated using student's t test and * and ** denotes p value of ≤0.05 and ≤0.01, respectively.