Fig. 2

ILC2 from 3xTg-AD mice exhibit intrinsic defects in producing IL-5. a 100 ILC2 were purified from the whole-brain tissue of 7-month-old female and male 3xTg-AD mice and control wild-type mice by fluorescence-activated cell sorting (FACS). Specifically, after liberase digestion, whole-brain tissue cells from the two mice of the group were pooled into one sample, and FACS were performed to sort ILC2. We pooled cells from two mice into one sample, in order to ensure that 100 ILC2 were obtained from each sample. One hundred ILC2 were plated into round-bottom 96-well plates and cultured with 20 ng/ml IL-2, IL-7, and IL-33 for 7 days. Growth of ILC2 was calculated as the number of cells that emerged after 7 days of culture per cell plated. Each culture well contained cells pooled from two different mice of the same group. Data are from 4 independent culture well per group, 2 independent experiments. b Expression of IL-5 was examined after 7 days of culture. Isotype was used as a negative control. c The mean fluorescence intensity (MFI) of IL-5 expression by cultured ILC2, after 7 days of culture. Data are from 5 independent culture well per group, 2 independent experiments. Sex effect: F [1, 16] = 0.57, p = 0.46; strain effect: F [1, 16] = 98.12, p < 0.01; interaction: F [1, 16] = 0.15, p = 0.71. d Concentrations of the indicated cytokines in the culture supernatant after 7 days of culture, measured by LegendPLex assays. Data are from 5 independent culture wells per group, 2 independent experiments. Sex effect: F [1, 16] = 0.19, p = 0.67; strain effect: F [1, 16] = 48.83, p < 0.01; interaction: F [1, 16] = 0.78, p = 0.39. U.D., undetected. Error bars = mean ± SEM. **p < 0.01; n.s., not significant