Figure 6

CCR7, FADD and TRAF6 are targets of miR-146a. Naïve CD8+ or CD8+/CD4+ T cells were nucleofected with either miR-146a mimic or scrambled control. After 48 hours culture in 10 ng/ml IL-7 cells were harvested and either (A) subjected to RT-PCR analysis to assess expression of putative targets of miR-146a namely CCR7, FADD and TRAF6 or (B) PCR was performed to analyse for the presence of various CD45 isoforms in cells following nucleofection. (A) Each bar represents the average of triplicates from three separate experiments and is expressed relative to the expression levels for each gene in cells nucleofected with the scrambled control (con) which is set to 1. Error bars represent standard deviation. A paired student’s t test was used to assess significance on non-normalised data * = p <0.05; ** = p <0.01. (B) Following CD45 specific cDNA synthesis and PCR, samples were visualised on an agarose gel. Lane 1 = scrambled control, lane 2 = miR-146a mimic. As a comparison CD45 isoforms present in either CD45RO + memory cells or ex vivo naïve were also assessed (lanes 3 and 5 respectively). Lane 4 = DNA ladder.