Figure 2.

IL-1 family gene expression is induced in islet immune cells in aging

(A) Representative blots detailing the sorting strategy used for isolation of beta cell, rest cell and immune cell fractions from 12- and 52-week-old RIP-EYFP reporter mice. Upper panel: gating for Forward-Side Scatter Area (FSC-A/SSC-A) for bulk cells selection; size and granularity of beta cells (green) and rest cells (red) in sorted fractions; gating of CD45-negative and CD45-positive cells, DAPI was used to remove dead cells; sorting gate for the immune cell fraction based on CD45 positivity and YFP negativity. Lower panel: from the CD45-negative cell gate (upper middle blot), beta cells (green) were gated as YFP-positive cells and rest cells (red) as YFP-negative cells; single cell sorting gates for beta cells (right blot) and rest cells (left blot).

(B–E) Relative gene expression in all FACS fractions expressed as fold of mean expression level of EYFP+ cells from 12-week-old beta cell reporter mice.

(F–K) Relative gene expression of purified (F and I) beta cell, (G and J) islet immune cell, and (H and K) rest cell fractions from 12-week-old and 52-week-old mice, normalized to mean expression level of 12-week-old mice; n = 7–10 beta cell fractions, n = 5–6 immune cell fractions, and n = 5–7 rest cell fraction from 7–10 mice per age group (in case of low yield, immune cells or rest cells from 2 age-matched mice were pooled).

Statistics: Student's t test; error bars represent SEM; ^★p < 0.05, ^★★p < 0.01, ^★★★p < 0.001.