Figure 4.

miR-466a inhibitor decreases pro-inflammatory and increases anti-inflammatory cells after coculture with alloantigen. Coculture of lymph node (LN) cells with alloantigen increases miR-466a-3p expression compared to LN cells cultured with syngeneic antigen at the indicated time points as determined by quantitative real-time-PCR (A). LN cells were administered alloantigen (50 µg/mL) for 10 days in complete media. Fresh media and miRNA inhibitors or controls were added every 3 days. Cells were harvested and stained for Tc1, Th1, Th17, and Treg cells. (B) Quantitation of flow cytometry plots. (C) Histogram of FoxP3 expression (bottom row), gated on CD4⁺, CD25⁺ dot plots (top row), and quantified in (D). Data are presented as mean ± SEM of two independent transfection experiments indicating six measurements. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001 by ANOVA with Tukey’s multiple comparisons test.