Fig. 6.
Expression of cytokine/chemokine mRNAs in the cell lysates of HeLa hVECs. HeLa cells were seeded at a density of 106/well in 24-well plates and treated with LPS (10 μg/ml for 6 h) or LPS-induced (10 μg/ml for 6 h) cells treated with RVFHbαP (70.45 μM for 1 h) or scrambled peptide (70.45 μM for 1 h). At the end of treatment, cells were collected, and lysates were prepared and analyzed for inflammatory mediators and GAPDH mRNA transcription by RT-PCR as detailed in Materials and Methods. (a) Representative image of RT-PCR analysis of cytokine/chemokine mRNA expression is shown. GAPDH blots confirmed roughly equivalent loading of RNA samples. Expression of cytokine genes was upregulated in LPS-induced cells and was significantly suppressed following the treatment of LPS-induced cells with RVFHbαP in respect to control values and is calculated as the mean ± SD of triplicate determinations performed on different days. (b) A quantitative assessment of the intensity of each band was determined by densitometry. Level of significance (**, P < 0.001 compared with the LPS- and LPS-nRVFHbαP-treated groups) was calculated by an ANOVA test followed by a Bonferroni analysis.