Fig. 2: BCL-2 expression defines p16+ cells with senescent characteristics.

a Schematic of multidimensional p16+ senescent cell analysis workflow; b UMAP visualization and FlowSOM clustering of pooled p16+ cells from young (n = 15) and old (n = 12) mice with c bar graphs indicating percent-of-whole cluster abundance changes. d FlowSOM clusters ranked by descending Ki67 mean expression plotted against p16 mean expression (Log2 Fold-Change). e Gating strategy for p16 + BCL2+ cells and f quantification of %Ki67+ and Ki67 mean expression alongside p16- and total p16+ cells. g Volcano plots of age-related changes in mean senescence marker expression within p16+ cells and quantification of % BCL-2+ cells and mean BCL-2 expression within all p16+ cells with age; h Gating strategy for “p16KB” cells; i Heatmap representation of protein expression between p16KB cells and non-senescent p16- cells, with asterisks indicating significance (*p < 0.05, **p < 0.01). Columns within each population represent a biological replicate (n = 12 Old). j Quantification of p16KB cells with age and numerical comparison to total p16+ cells (n = 15 young and n = 12 old biologically independent animals). Schematic in (a) was generated using BioRender. Box plots show median and interquartile range with error bars representing minimum and maximum values. (f, g [volcano plot]) Multiple two-sided t tests with Holm-Sidak Correction. (g, i, j) Two-sided Mann–Whitney test. Source data are provided as a Source Data file.