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. Author manuscript; available in PMC: 2017 Jul 19.
Published in final edited form as: Immunity. 2016 Jul 19;45(1):83–93. doi: 10.1016/j.immuni.2016.06.022

Figure 3. Identification of miR-17 as an Eos 3′UTR targeting miRNA.

Figure 3

(A) Expression of Eos mRNA in Treg cells after stimulation. CD4+ Foxp3GFP+ Treg cells were isolated by FACS and incubated with anti-CD3/CD28 antibodies for the indicated times and Eos mRNA levels were determined by RT-PCR. (B) The psBZN - iCHECK2 reporter vector (Bozeman, Montana, USA) shown was used to assess the ability of miRNAs to target the Eos 3′UTR. (C) Luciferase reporters containing the WT Eos 3′UTR or mutant Eos 3′ (MU). miR-17-92, miR-17-19a, and miR-17 were screen for inhibition of Eos 3′UTR luciferase activity relative to mock treatment. Rluc =Renilla luciferase; Luc = firefly luciferase; and pA =polyadenylation signal. The pcDNA3 vector was used to transiently express miRNAs. Eos 3′ MU contained mutations at miR-17 and miR-92 sites and served as a negative control for miRNA inhibition.