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. 2018 Oct 9;9:2232. doi: 10.3389/fimmu.2018.02232

Figure 5.

Figure 5

miR-155 is required for ILC2 expansion in vitro but not IL-13 production. ILC2 were isolated from MLN of IL-2/anti-IL-2/IL-25 treated WT or miR155−/− mice. ILC2 were either left untransduced or transduced with a retrovirus over-expressing the miR-155 sequence to restore miR-155 signaling. Cells were cultured for 6 days in the presence of IL-7 and IL-33 (both at 10 ng/ml). (A) Daily ILC2 counts. (B) Intracellular IL-13 staining of WT or miR155−/− ILC2 from 5-day cultures in (A) analyzed by flow cytometry. (C) Secreted IL-13 was quantified in supernatants from the cultures of WT and miR155−/− ILC2 using ELISA (data corrected for cell counts). (D) Secreted IL-13 was quantified in supernatants from WT ILC2 treated with negative control siRNA or miR-155 knockdown siRNA harvested after 2 days using ELISA (data corrected for cell counts). (E) Intracellular IL-13 expression in ILC2 isolated from the MLN of IL-33-treated lethally-irradiated B6SJL mice that had received mixed BM from Roraflox/flox x Il7raCre mice with WT or miR155−/− BM. Cells were treated with PMA/ionomycin in the presence of protein transport inhibitor. Data are representative of at least two independent experiments. Two-way ANOVA with Sidak's multiple comparisons test (A), #p ≤ 0.05 compared to miR155−/−, §p ≤ 0.05 compared to control transduced miR-155−/− ILC2.