Figure 6

Ectopic expression of miR-17-92 cluster members in the human Jurkat T cell line confers increased IL-2 production and resistance to AICD. Jurkat cells were transduced by either one of the following pseudo typed lentivirus vectors: 1) control vector encoding GFP; 2) the 17-92-1 expression vector encoding miR-17 18 and 19a, or 3) the 17-92-2 expression vector encoding miR 20, 19b-1, and 92a-1. (A), Transduced Jurkat cells (5 × 10⁴) in the triplicate wells were stimulated with PMA (10 ng/ml) and ionomycin (500 nM) for overnight and supernatant was harvested and tested for the presence of IL-2 by specific ELISA. The figure shows mean values and standard deviations of the amount of IL-2 released from each group. Statistical analysis was carried out using the student t test, and significant (p < .005) increase of IL-2 production was confirmed in both 17-92-1 and the 17-92-2 transduced groups compared with the control group. (B), Transduced Jurkat cells were treated with the AICD inducing condition (10 μg/ml anti-CD3 mAb) or in complete media (No Tx) for 24 hrs. Then, the relative numbers of viable cells were evaluated by 4 hour WST-1 assays. The figure shows mean values and standard deviations of 8 wells/group each containing 5 × 10⁵ cells. For each group, the relative OD readings at 450 nm of AICD-treated cells compared with control Jurkat cells without AICD-treatment is indicated. * indicates p < 0.05 between the two groups using student t test.