A. QPCR analysis of expression of miRs – 21, 22 and 146b following ectopic expression of Bcl6 in naïve T cells. Sorted RV+ T cells were re-stimulated for 6 hrs to assess its effect on miR expression. Data are averaged from at least 3 different experiments.
B. Luciferase activity in Jurkat T cells co-transfected with full-length or SB1 and SB2 mutated miR-21 promoter driven luciferase reporters and expression constructs for CXN and CXN-Bcl6. Cells were stimulated with PMA and Ionomycin for 24 hrs prior to harvest and luciferase measurement. Results are averaged from 5 independent experiments, where the basal luciferase activity of each promoter construct was set to 100%.
C. QPCR analysis of miR-21 expression following culture of WT, Bcl6−/− and Stat3−/− naïve T cells (CD4+CD62L+) in response to increasing doses of IL-6 (0, 0.1, 1, 10 ng/ml) for 24 hrs, with expression normalized to U6. Data representative of 2 independent experiments.
(A–B) *p<0.05, **p<0.01 (Student’s t-test) (error bars=s.e.m.)