Fig. 3.
a: Meth modulates the gene expression of TJ proteins in BMVEC. BMVEC (1×106 cells/ml) were treated in vitro with Meth (10 nM) ± gp120 (50 ng) for 48 h. RNA was extracted, reverse transcribed, cDNA amplified and the ZO-1, JAM-2, Claudin-3, Claudin-5 and Occludin gene expressions were determined by real time quantitative PCR. Relative expression of mRNA species was calculated using the comparative CT method. Data are the mean±SD of 3 separate experiments done in triplicate. Statistical significance was determined using ANOVA based on comparison with the untreated control. Our results show that the most significant differences in TJ expression were observed when the BMVEC were treated with a combination of Meth and gp120. b: An amplification plot from a representative real time quantitative PCR run. The line across the amplification plot is called the threshold, which determines the CT value and is selected where all data are undergoing exponential amplification. The threshold is a numerical value assigned automatically by the software for each run above the calculated baseline.