Figure 2: Post-transcriptional Dicer dependent accumulation of miR-146a.

(a) Relative expression of pri-miR-146a in naive T cells sorted from Roquin^+/+ and Roquin^san/san mice measured by qRT–PCR using three different sets of primers shown as (i), (ii) and (iii) in the schematic. NS, not significant (P<0.05, U-test). (b) The levels of pre-miR-146a in Roquin^+/+ and Roquin^san/san naive T cells measured by qRT–PCR. (c) Northern blots showing precursor and mature miRNA transcripts (arrows) of miR-146a and miR-150 in naive T cells of Roquin^san/san and Roquin^+/+ mice. U6 snRNA was used as a loading control and is shown below the miR-146a northern blot. Weaker less abundant bands are miRNA decay products. M, marker. (d) Relative expression of miR-146a in Dicer-floxed-Rosa26-Cre-ER MEFs (Dicer1^−/−) and CD4-CRE (wt) control MEFs assessed by qRT–PCR. MEFs were retrovirally transduced with Roquin^wt and Roquin^san containing a GFP reporter or with an empty GFP vector control. After 2 days, MEFs were treated with tamoxifen to delete Dicer. GFP-positive cells were sorted 7 days later. Each dot represents individual mice (a,b) or technical replicates (d) and the bars represent the median value in each group. U-test: *P<0.05, **P<0.005. Results are representative of two independent experiments.