Fig. 2

Characterization of mouse islet immune cells during ageing. Islets were isolated from C57Bl/6 J male mice aged 3, 6, 12 and 24 months for analysis by flow cytometry. a Schematic of flow cytometry protocol; islets from a minimum of 5 mice were pooled per sample to obtain sufficient dispersed islet CD45+ cells for analysis. Each pooled sample of ≥5 mice is considered one individual biological sample (n = 1). b Cells were gated on forward-scatter (FSC), side-scatter (SSC), viability (FVD-), CD45+, and subsequently CD19 or CD5. Data are from a representative 6-month old mouse islet sample. c-d CD45+ and CD19+ cells expressed as a percent of total viable islet cells. e Representative plots of SSC^lowCD5+ populations from 3-month and 24-month old mouse islets. f Cells gated on SSC^lowCD5+ are positive for CD3+, box represents CD3 + CD5+ cells. g-j T cells expressed as a percent of total viable islet cells (G-H) or CD45+ islet cells (i-j). Linear regression (h,j) analysis shows 95% confidence interval in blue and Pearson correlation coefficient, connecting lines between data points are from a given cohort of mice run in one independent experiment. Data represent independent biological samples, each run as an independent experiment, with mean ± SD overlaid (c,d,g-j), and were analyzed by one way ANOVA with Dunnett’s T3 multiple comparisons test (c,g,i) or Kruskal-Wallis test with Dunn’s multiple comparisons test (d). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Numbers in FACS plots represent the percent of cells in each selection as a function of the parent population