Contribution of Kras ^G12D -mutant HSC is delayed by osteopontin deficiency. (A) Feature plots showing the expression of Tnf (encoding TNF-α) or Spp1 (encoding osteopontin) in individual cells on the KNetL plot (Fig. 4). The Kras^G12D-specific MPP cluster is highlighted. (B–E) Contribution of Kras^G12D-mutant HSC with or without osteopontin. HSC-specific Cre reporter mice (Pdzk1ip1-CreER R26^Tom/Tom) that carried WT Kras allele, a conditional mutant (Kras^G12D) Kras allele alone, or Kras^G12D allele with a germline deletion of Spp1 (Spp1^−/−) were administered a single dose of tamoxifen to induce simultaneous Tom expression and Kras mutation in HSC, and analyzed at the 3-wk endpoint. Shown is the fraction of Tom⁺ cells within mature BM cell types (B), subsets of B cells in the BM (C), and subsets of thymocytes (D) and stem/progenitors in the BM (E). Symbols represent individual mice; bars represent the mean ± SD; data are pooled from two independent experiments. Statistical significance was estimated by the Mann–Whitney test. *P < 0.05, **P < 0.01, ***P < 0.001.