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. Author manuscript; available in PMC: 2013 Jun 1.
Published in final edited form as: Genes Immun. 2012 Sep 20;13(8):605–620. doi: 10.1038/gene.2012.39

Figure 6. PAR-1 inhibitor SCH79797 regulates the expression of MCP-1, MMP-1, and MMP-9 in THP-1 monocytic cells stimulated by sonicated H37Rv M. tuberculosis and THP-1 cells stimulated by sonicated H37Rv M. tuberculosis in the presence of exogenous purified human MMP-1.

Figure 6

Panel 1, we measured the relative changes in MCP-1, MMP-1, MMP-9, and TIMP gene expression by real-time PCR. Data are presented as the fold change in gene expression normalized to the endogenous reference gene PDHB and relative to untreated controls. The effect of 50 nM concentration of SCH79797 PAR-1 inhibiting compound was assessed following 24 hr in vitro stimulation of THP-1 cells with 5 μg/ml sonicated H37Rv M. tuberculosis. Cultures proceeded in 500 μl incomplete RPMI. PAR-1 SCH79797 inhibiting compound was diluted with DMSO and dispensed in 5 μl volume to produce a final culture concentration of 0.01% DMSO. We also added 5 μl of DMSO to control cultures. The results presented are from six independent experiments showing the mean and standard deviations. We consistently observed significant differences in the mean values across variables (Kruskal-Wallis p < 0.001) when testing MCP-1, MMP-1 and MMP-9. We show corrected p-values obtained from student t-tests. Of note, levels of specific mRNAs from non-stimulated cells were not significantly different from those obtained from cultures that proceeded with the PAR-1 inhibitor alone. Panel 2, the effect of 1 nM exogenous human purified MMP-1 on THP-1 cells exposed to 5 μg/ml sonicated H37Rv M. tuberculosis was assessed. We also assessed the effect of 1 nM exogenous human purified MMP-1 on THP-1 cells exposed to 5 μg/ml sonicated H37Rv M. tuberculosis in the presence or absence of 50 nM concentration of SCH79797 PAR-1 inhibiting compound. We measured the relative changes in MCP-1, MMP-1, and MMP-9 gene expression by real-time PCR as explained above. Cultures proceeded in 500 μl of incomplete RPMI. The results presented are from three independent experiments showing the mean and standard deviations. We consistently observed significant differences in the mean values across variables (Kruskal-Wallis p < 0.001) when testing MCP-1, MMP-1 and MMP-9. We show corrected p-values obtained from student t-tests. The inhibitor’s concentration was selected from dose response-experiments.