TCR signaling pathways affecting miR-146a expression. Jurkat cells were treated for 30 minutes with the JNK inhibitor SP600125, the MEK/ERK inhibitor PD98059, or CsA. Thereafter, cells were stimulated for 24 hours with anti-CD3/CD28 Abs. (A) Relative IL-2 mRNA levels were measured by qRT-PCR using GAPDH levels to normalize expression. Relative IL-2 mRNA levels in stimulated cells are compared with, which are set to 1. Each bar is the mean of 3 independent experiments. (B) MiR-146a levels were measured by qRT-PCR and U6 levels were used to normalize expression. Relative miR-146a expression in stimulated cells is compared with that in nonstimulated cells, which set to 1. Each bar is the mean of 3 independent experiments.