Overexpression of miR-146a in CAL-1 cells blocks TLR-induced NF-κB activation. (A) Flow cytometric analysis of phospho-p65 levels after intracellular staining of CAL-1 cells. CAL-1 cells were transduced with miRNA-146a or with control hTR-expressing vectors, which also drive expression of the marker GFP. Levels of phosphorylation of the NF-κB subunit p65 were measured in GFP-sorted transduced cells after activation for 15 minutes with 10 μg/mL CpG-B (dark gray line), 10 μg/mL R848 (black line), or medium as control (light gray line). RCN, relative cell number. (B) Statistical analysis of phospo-p65 levels was assessed by plotting the difference in mean fluorescence intensity (ΔMFI) between activated cells and medium-cultured cells of 4 different experiments. Data are normalized to phospho-p65 levels in activated hTR-transduced cells, which was set to 1. *P = .014; **P = .004.