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. 2014 Dec 11;290(5):2831–2841. doi: 10.1074/jbc.M114.591420

FIGURE 2.

FIGURE 2.

miR-146 modulates DC apoptosis. imDCs at day 4 were transfected with miR-146a and/or miR-146b and scrambled control (Ctrl) inhibitors (A–C) or mimics (E–G) at a final concentration of 30 nm for 40 h. A Cy3-labeled negative control was used to monitor the transfection efficiency (close to 80%) of these miRNA inhibitors or mimics. A and E, the relative expression of miR-146a and miR-146b was quantified by real-time PCR. C and G, the percentages of annexin V+ cells (including annexin V+/propidium iodide− (PI) and annexin V+/propidium iodide+) were determined as in B or F. imDCs at day 6 were transfected with miR-146a and/or miR-146b and scrambled control inhibitors (D) or mimics (H), followed by the maturation mixture for 40 h, and cells were then analyzed for apoptosis as described above. Caspase-3/7 activity was measured with the Caspase-Glo 3/7 assay after transfection of miR-146a and/or miR-146b and scrambled control inhibitors or mimics for 40 h in imDCs (I) and mDCs (J). Data shown are mean percentages ± S.E. of eight (C), four (A and E), six (D, G, and H), or two (I and J) independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.005 (by paired t test).