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. Author manuscript; available in PMC: 2022 Mar 1.
Published in final edited form as: Nature. 2021 Aug 11;597(7875):256–262. doi: 10.1038/s41586-021-03795-7

Fig. 2 ∣. The SSC lineage contributes to age-related skewing of the haematopoietic lineage.

Fig. 2 ∣

a, Schematic representation of parabiosis experiments with isochronic 2-month-old (isochronic young; IY) and 24-month old (isochronic aged; IA) pairs as well as heterochronic pairs with one 2-month-old (heterochronic young; HY) and one 24-month-old (heterochronic aged; HA) mouse. b, BMD at four weeks of parabiosis (IY, n = 6; HY, HA, IA, n = 4 per group). c, Frequency of SSCs and BCSPs at four weeks of parabiosis. d, BMD at day 10 after fracture of parabiosed mice (IY, n = 6; HY, n = 3; HA, n = 3; IA, n = 3). e, In vitro osteogenesis of SSCs from parabiosed mice at day 10 after fracture showing representative staining (top) and quantification (bottom). OD450 nm, optical density at 450 nm. f, In vitro chondrogenesis of the same SSCs (n = 3 per group). g, Schematic for transplantation of GFP-labelled HSCs from the bone marrow of 2-month-old mice into lethally irradiated 2-month-old or 24-month-old mice. h, Peripheral blood analysis at 6 weeks (wks) and 12 weeks after haematopoietic reconstitution for lymphoid (B and T cells) and myeloid (GR1+) fractions (2-mo, n = 6; 24-mo, n = 5). i, Expression of CD150 (SLAM) in donor-derived GFP+LIN−cKIT+SCA1+FLT3−CD34− bone marrow HSCs. j, Bone marrow analysis of donor-derived (GFP+) haematopoietic cell populations at 12 weeks by flow cytometry (2-mo, n = 6; 24-mo, n = 5). CLP, common lymphoid progenitor; GMP, granulocyte–monocyte progenitor. k, Schematic of SSC–HSC co-culture experiments. l, Peripheral blood analysis after haematopoietic reconstitution with co-cultured haematopoietic cells. m, Bone marrow analysis of co-cultured donor-derived (GFP+) haematopoietic cell populations at 12 weeks (no SSCs, n = 3; 2-mo, n = 4; 24-mo, n = 3). Data are mean + s.e.m. Statistical testing in b–f, m by one-way ANOVA analyses with Tukey’s post-hoc test for all comparisons. j, Two-sided Student’s t-test adjusted for non-normality (Mann–Whitney test) where appropriate. h, l, Two-way ANOVA with Bonferroni post-hoc test. For exact P values, see Source Data. Scale bars, 50 μm.