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. 2010 Jun;51(6):2976–2985. doi: 10.1167/iovs.09-4874

Figure 8.

Figure 8.

Role of PAI-1 and IRAK1 on the effects of miR-146a on the production of iROS and BrdU incorporation. Presenescent HTM cells at p11 were cotransfected with 2 μg of either a negative control plasmid (pCon), a plasmid expressing PAI-1 (pPAI-1), or a plasmid expressing IRAK1 (pIRAK1, which lacked the 3′UTR that contains the target site for miR-146a) and 120 pmol of either a miR-146a mimic (146aM) or a scrambled miRNA used as a negative control (ConM). Three days after transfection, production of iROS was determined by DCFH oxidation, and cell proliferation was measured by BrdU incorporation. (A) Comparison cells cotransfected with pPAI-1/ConM with those cotransfected with pCon/ConM showed that overexpression of PAI-1 resulted in an increase of almost twofold in the generation of iROS. However, a comparison between cells cotransfected with pPAI-1/146aM or pPAI-1/ConM showed that miR-146 decreased the production of iROS in cells overexpressing PAI-1. In contrast, expression of IRAK1 lacking the 3′UTR prevented completely the decrease in iROS production mediated by miR-146a. (B) Overexpression of either PAI-1 or IRAK1 lacking the 3′UTR prevented the increase in cell proliferation induced by miR-146a. Data represent the mean percentage changes ± SD, n = 3–4. *#P < 0.05, compared with ConM+pCon by Mann–Whitney U test.