Hematopoietic Tet2^+/− clonal expansion rate increases in aged mice and associates with increased IL-1 BM levels. (A) Experimental design. (B) Longitudinal quantification of the percentage of PB CD45⁺ WT ZsG⁺ (n = 6; purple line) and CD45⁺Tet2^+/− ZsG⁺ (n = 7; red line) cells over 1 year after TAM induction. (C) Representative fluorescence-activated cell sorting plot of ZsG expression on PB CD45⁺ cells in 12-month-old WT and Tet2^+/− mice. (D) Percentage of WT (n = 6) or Tet2^+/− (n = 7) ZsG⁺ cells in PB T cells, B cells, myeloid cells, erythrocytes, and platelets. (E) Percentage of WT ZsG⁺ (n = 6) and Tet2^+/− ZsG⁺ (n = 7) on indicated BM populations. (F) Monthly expansion rates of PB CD45⁺ WT ZsG⁺ (n = 6) and CD45⁺Tet2^+/− ZsG⁺ (n = 7) populations at indicated time intervals. (G) Gene expression levels of indicated genes in WT ZsG⁺ (n = 3) and Tet2^+/− ZsG⁺ (n = 3) in total BM white blood cells, 1 year after TAM induction. (H) IL-1α and IL-1β protein levels in BM lysates of WT (n = 6) or Tet2^+/− (n = 7) mice, 1 year after TAM induction. (I) Correlation between BM IL-1α/IL-1β levels and the percentage of BM Tet2^+/− ZsG⁺ cells (n = 11). Pearson correlation coefficient (r) and P values are indicated. Dark red dots indicate mice from Figure 1B; light red dots indicate additional mice with low Tet2^+/− ZsG⁺ fractions. (J) Experimental design. (K) Fraction of T cells, B cells, or myeloid cells in WT (ZsG^–; n = 3) or Tet2^+/− (ZsG⁺; n = 3) BM CD45⁺ cells. L. IL-1α (left) and IL-1β (right) protein levels in the conditioned media (CM) of sorted WT and Tet2^+/− T cells, B cells, and myeloid cells without or with LPS exposure (n = 3). Data are a pool of at least 2 independent experiments for all graphs, except J through L. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 by unpaired t-test (B, E–H, and K) or by 1-way analysis of variance with Tukey correction (L). Error bars represent standard error of the mean. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ns, not significant.