Fig. 6: Effect of CALR del52 and CALR ins5 on the HSCs.

a Percentages of HSCs (SLAM, Lin⁻c-Kit⁺Sca-⁺CD150⁻CD48⁺) in BM and spleen of ≥10 months post-tamoxifen mice were expressed as means ± SEM (n = 5 independent mice of +/+ (white), +/del52 (orange) and +/ins5 (blue) genotypes; n = 4 del52/del52 (red) mice and for ins5/ins5 (green) genotype n = 4 for BM and n = 3 for spleen analyses). Significance was determined using a two-sided parametric t-test: in BM, *p = 0.047; **p = 0.0012 (+/+ vs. ins5/ins5), 0.0016 (+/del52 vs. del52/del52), 0.0034 (del52/del52 vs. ins5/ins5); ***p = 0.0004 except 0.0009 for +/ins5 vs. ins5/ins5 and in spleen, *p = 0.0157 (+/del52 vs. del52/del52), 0.0131 (+/+ vs. ins5/ins5); **p = 0.0071 (+/+ vs. del52/del52), 0.0094 (del52/del52 vs. +/ins5), 0.0334 (+/ins5 vs. ins5/ins5). b To investigate the effect of CALR del52 and CALR ins5 on HSCs we engrafted different ratios of homozygous del52/del52 (yellow, pink, red, brown and black squares and solid lines) and ins5/ins5 (turquoise, blue, green, dark blue and black squares with solid lines) CD45.2⁺ KI BM cells with wild-type (+/+) CD45.2⁺ GFP⁺ BM cells in CD45.1⁺ lethally irradiated mice. Four weeks after transplantation, KI allele expression was induced by tamoxifen and c platelet and d WBC counts were measured. Data are the means ± SEM, del52/del52 platelets: n = 4, 4, 2, 4, 4, 3 for 0%, n = 4, 4, 3, 3, 3, 3 for 10%, n = 5, 4, 5, 4, 4, 4 for 25%, n = 5, 5, 4, 5, 5, 4 for 50%, n = 3, 4, 4, 3, 4, 3 for 75% and n = 5, 5, 5, 3, 5, 2 for 100%; ins5/ins5 platelets: n = 3, 3, 3, 1, 3, 3 for 0%, n = 5, 4, 3, 4, 2, 4 for 10%, n = 5, 5, 5, 4, 2, 3 for 25%, n = 5, 5, 3, 4, 3, 5 for 50%, n = 5, 5, 3, 4, 3, 5 for 75% and n = 5, 5, 3, 4, 4, 5 for 100%; del52/del52 WBC: n = 4, 4, 3, 4, 4, 4 for 0%, n = 4 for 10%, n = 5, 4, 5, 5, 4, 5 for 25%, n = 5, 5, 5, 5, 5, 4 for 50%, n = 4, 4, 4, 3, 4, 4 for 75% and n = 5, 5, 5, 4, 5 for 100%; ins5/ins5 WBC: n = 3, 3, 3, 2, 3, 3 for 0%, n = 5, 5, 4, 4, 5, 5 for 10%, n = 5, 5, 5, 5, 3, 4 for 25%, n = 5, 5, 5, 5, 3, 5 for 50%, n = 5, 5, 3, 5, 4, 5 for 75% and n = 5, 4, 5, 5, 5, 5 for 100%. N represents individual mice and varies depending on time of sampling during the course of the experiment and on the group defined as the percentage of engrafted homozygous cells, as indicated in the Source data file. Significance compared to the 0% condition (open circles with connecting black dashed line) was determined using ANOVA with Dunnett’s multiple comparison test and a nonparametric Kruskal–Wallis’s multiple comparison test for ins5/ins5 WBC. In c, *p = 0.0172 (100%), 0.0256 (75%); ***p = 0.0001 (100% and 75%) and in d, *p = 0.0422 (100%), 0.0128 (75%); ns not significant. Source data are provided as a Source data file.