Fig. 2
From: Roquin targets mRNAs in a 3′-UTR-specific manner by different modes of regulation
Nfkbid is strongly regulated by Roquin on the mRNA and protein level. a–c Immunoblot (a), flow cytometry (b) and quantitative RT-PCR (c) analysis of previously described Roquin targets in CD4+ T cells derived from Rc3h1-2fl/fl; Cd4-Cre-ERT2 (iDKO) mice treated in vitro with 4′ OH-tamoxifen. As wild-type (WT) controls either T cells from Rc3h1-2+/+; Cd4-Cre-ERT2 (WT) mice were treated in vitro with 4′ OH-tamoxifen or T cells from Rc3h1-2fl/fl; Cd4-Cre-ERT2 mice were left untreated. d Immunoblot analysis of Roquin-1 and Roquin-2 proteins before and after 4’ OH-tamoxifen treatment of MEF cells that contained Rc3h1 and Rc3h2 alleles with loxP-flanked exons and a Cre-ERT2 transgene. e Flow cytometry analysis of GFP in Rc3h1–2fl/fl, Cre-ERT2 MEF cells retrovirally transduced with a reporter construct encoding GFP and human ICOS coding sequence (ICOS–CDS) followed by the full length Nfkbid 3′-UTR (1–559) harboring two CDE, one ADE and one LBE element or truncated versions (scheme) of the 3'-UTR, either treated with 4′ OH-tamoxifen (iDKO) or left untreated (WT). Statistical significance was calculated by unpaired two-tailed Student’s t test (b, c); ns = not significant; ***p < 0.001, **p < 0.01. Error bars indicate mean ± SEM. Data are representative of three (a, b, d, e) and seven (c) independent experiments
