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. 2019 Sep 18;10:2051. doi: 10.3389/fimmu.2019.02051

Figure 3.

Figure 3

Butyrate regulates GATA3 expression and ILC2 metabolism. Activated ILC2s were FACS purified from lungs after 3 days IL-33 administration i.n. and cultured ex-vivo for 24 h in the presence of 200 uM Sodium Butyrate or Sodium Chloride as control for mRNA expression (A) or incubated with indicated SCFA sodium salt at 200 uM for 2–3 days (B–D). (A) mRNA quantification of immune-related genes by Nanostring panel shown as heatmap of absolute expression counts. (B) Surface expression of ST2 and Intracellular expression of GATA3 in response to indicate SCFA. (C) Oxygen consumption measure (OCR) from Seahorse mito stress test 48 h post butyrate treatment. (D–F) Activated ILC2s were cultured ex-vivo for 2 days in the presence of 1 uM GATA3 Morpholino (GATA3 MO) or 5' mismatch control (CTRL MO) followed by another 2 days of Acetate or Butyrate treatment in presence of 10 ng/mL rmIL-33. (D) GATA3 expression over isotype. (E) IL-5 and IL-13 cytokine secretion in supernatant after acetate or butyrate addition. (F) Seahorse mito stress test assay after GATA3 knockdown ILC2 showing OCR and ECAR response. Data are graphed as duplicate-triplicate wells +-SEM, representative of 2–3 independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001.