FIGURE 2. c-Myc is required for ILC2 activation in vitro.

(A) ILC2/b6 cells were transduced with lenti-CRSIPRv2-GFP knockout constructs targeting c-Myc or non-target control (NTC). GFP⁺ cells were sorted on 10 days post transduction, and expression of c-Myc was examined by western blot. (B) Primary mouse ILC2 were isolated from the lungs of naïve mice, cultured with IL-33, IL-7 and IL-2, and transduced with lenti-CRSIPRv2-GFP knockout constructs targeting NTC or c-Myc. Growth of GFP⁺ cells was calculated as fold change in cell number over 5 days of culture. (C) Expression of IL-5 and IL-13 by GFP⁺ ILC2 were examined at 10 days after lentiviral transduction. (D) ILC2/b6 cells were transduced with lenti-CRSIPRv2-GFP knockout constructs targeting NTC or c-Myc. Genome-wide microarray was performed with sorted GFP⁺ cells at 10 days post transduction, and gene set enrichment analyses were performed. (E) Heatmap of representative genes by microarray analysis. (F) Primary mouse ILC2 were isolated from the lungs of naïve mice, cultured with IL-33, IL-7 and IL-2, and transduced with lenti-CRSIPRv2-GFP knockout constructs targeting NTC or c-Myc. mRNA level of Stat6 in GFP⁺ ILC2 was measured by qPCR analysis at 10 days post transduction. (G) CHIP analysis was performed in ILC2/b6 cells. DNA region lacking c-Myc binding site was used as a negative control. Data are representative of three independent experiments (A), or are from three or four mice per group, representative of two independent experiments (B and C), or are from three biological replicate samples (D and E), or are from 3-4 samples, representative of two to four independent experiments (F and G). *P < 0.05.