Figure 2

15d-PGJ2 inhibits mRNA and protein expression of endothelial CAMs. bEND.3 cells were pretreated with 15d-PGJ2 (5–20 μM) for 30 min followed by stimulation with TNFα (50 ng/ml) for 2 h. Cells were harvested in Trizol reagent for RNA isolation and cDNA synthesis. RT-PCR analysis was done for ICAM-1 (A), VCAM-1 (B) and E-selectin (C). Results were calculated as mean ± SD for 3 independent experiments. Samples were examined in triplicates. &&& p < 0.001 compared with control (untreated and unstimulated cells) and !!! p < 0.001 as compared to TNFα treatment. For the quantitation of expression of surface CAMs, bEND.3 cells were treated with TNFα (50 ng/ml) in the presence or absence of 15d-PGJ2 (5–20 μM) for 6 h followed by flow cytometry analysis (D) (n = 2).