Extended Data Figure 7: Additional analyses of old Gfp–Lc3 HSCs.
From: Autophagy maintains the metabolism and function of young and old stem cells
a, Lineage distribution in peripheral blood (left) and frequency of the indicated populations (right) in young and old Gfp–Lc3 mice (related to Fig. 5a, b). b, pS6 levels in yHSCs and oHSCs. c, d, Representative FACS plots and quantification of Cyto-ID dye levels in (c) HSCs cultured for 3 h with or without cytokines, and (d) yHSCs and oHSCs. e, Representative FACS plots of autophagy low (ATlo) and autophagy high (AThi) oHSC subsets. f, Representative examples of three independent experiments showing immunofluorescence staining of FOXO3a in ATlo and AThi oHSCs. Scale bar, 10 μm. g, Representative electron micrographs of oHSCs with or without autophagosomes (AP) showing vesicles (top) and endoplasmic reticulum/Golgi (bottom) compartments. Scale bar, 1 μm. h, Representative immunofluorescence staining and quantification of TOM20 in ATlo and AThi oHSCs. Scale bar, 10 μm. i, j, Characteristics of ATlo and AThi oHSCs: (i) TMRE levels, (j) ATP levels, and (k) ROS levels measured by Mitosox Red (MSR) staining. l, Expansion of ATlo and AThi oHSCs in self-renewal culture conditions. m, Colony formation in methylcellulose from ATlo and AThi oHSCs. Results are expressed as percentage of 100 plated cells. Data are mean ± s.d., and are expressed relative to +cyt HSC (c), yHSC (b, d), or ATlo oHSC (h–j, l) levels. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.
