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. Author manuscript; available in PMC: 2014 Jun 15.
Published in final edited form as: J Immunol. 2013 May 17;190(12):5972–5980. doi: 10.4049/jimmunol.1300351

Figure 4. Ets1 is a functionally relevant target of miR-155 in Th17 cells.

Figure 4

(A) Schematic of experimental design. (B) miR-155+/+ or miR-155−/− IL-17F RFP+/− CD4+ T cells were isolated and infected with a control (NC) or Ets1 shRNA expressing retrovirus. After culturing under Th17 cell-skewing conditions for 72 hrs, RNA or protein was extracted and expression of the indicated Th17-related genes were analyzed by qPCR (Upper panel) or Ets1 by Western blotting (Lower panel). (C) miR-155+/+ or miR-155−/−RFP+GFP+ T cells were sorted by FACS and subjected to intracellular staining for IL-17A. (D) Graph of the average relative percentage of IL-17A+ cells in sorted miR-155+/+ or miR-155−/− RFP+GFP+ cells transduced with the control or Ets1 shRNA expressing retroviral vector (n=4, 2 independent experiments). (E) Expression of Th17 cell effector genes in sorted miR-155−/− RFP+GFP+ cells transduced with the control or Ets1 shRNA expressing retroviral vector were analyzed by qPCR (n=4). Error bars represent +/− SEM. * denotes a p value of <0.05.