MiR-146a overexpression induces apoptosis of CAL-1 cells. (A) CAL-1 cells were transduced with hTR control RNA (open symbols) or miR-146a (closed symbols) and cultured in medium. GFP expression was determined at the indicated days by flow cytometry. Shown are the absolute numbers of GFP⁺ CAL-1 cells. (B) GFP⁺ cells were sorted and cultured for 2 days. Cells were stained with Annexin V and 7-AAD to assess for apoptosis. Numbers in the dot plots represent percentages of cells that fall within the indicated gates. (C) The percentages of apoptotic cells were determined as in panel B. Shown are mean percentages ± SD of 3 independent experiments (*P < .05). (D) QPCR analysis of BCL2-A1 mRNA levels in CAL-1 cells transduced with hTR control or with miR-146a. Expression levels in hTR-transduced CAL-1 cells were set to 1. Shown are mean values ± SD of an analysis of QPCR done in triplicate. This is a representative experiment of 4. **P < .01.