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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 5. miR-17 modulates Treg cell suppressive activity.

Figure 5

(A) The effect of miR-17 overexpression on the in vitro function of Treg cells was explored by transducing them with either a bicistronic retrovirus expressing miR-17 or the GFP-containing empty vector (EV) prior to co-culture with CFSE labeled CD4+ naïve T cells from CD45.1+ mice at the indicated ratios in the presence of anti-CD3/CD28 beads for 4 days. CD45.1+ T cells were analyzed by flow cytometry for CFSE dilution (blue line = Tnaive only; red line= Treg cells and Tnaive). (B) The in vitro suppressive capacities of Treg cells transduced with either the antisense construct, miRZip17 or a control vector were determined as in (A). (C–F) The impact of miR-17 on Foxp3 induction and pro-inflammatory cytokine expression by iTreg cells. Naïve T cells from WT mice were transduced as in (A), followed by in vitro Treg skewing for 72h. Intracellular levels of Foxp3 (C) or Foxp3 and IFNγ were determined by flow cytometry after restimulation with PMA and ionomycin (D). Data are representative of 4 independent experiments. (E, F) qRT-PCR analysis of IFNγ and Eos mRNA in Treg cells from (C). Shown are mean +/− SEM from three independent experiments.