Extended Data Figure 4: Comparison between aHSCs, Atg12cKO HSCs and oHSCs.
From: Autophagy maintains the metabolism and function of young and old stem cells
a, Representative FACS plot of pS6 levels in HSC cultured for 1 h (hr) with or without cytokines (±cyt). b, Representative FACS plot of GFP–LC3 in freshly isolated HSCs (t = 0) and HSCs cultured for 6 h with or without cytokines. c, Inactivation of autophagy in +cyt measured by p62 levels. d, Drug modulation of autophagy levels in Gfp–Lc3 HSCs cultured for 3 h with or without cytokines. Rap, rapamycin; I128, Ink128; CC, compound C. Results are expressed as percentage GFP–LC3 levels upon 3 h culture in +cyt conditions. e, Cell size measured by forward scatter (FSC) in control and cKO HSCs, and yHSCs and oHSCs. f, g, Glycolysis activity measured by extracellular acidification rate (ECAR) in (f) control and cKO LSKs, and (g) yHSCs and oHSCs. Oligo, oligomycin; 2-DG, 2-deoxy-d-glucose. h, Mitochondria parameters in yHSCs and oHSCs. i, ROS levels in aHSCs. j, Representative FACS plot and quantification of ROS levels in control and cKO HSCs. k, Representative examples of three independent experiments showing immunofluorescence co-staining of γH2AX with fibrillarin (FBN), 53BP1, and RAD51 in control and cKO HSCs. Sscale bar, 10 μm. Data are mean ± s.d. except for line graphs, and are expressed relative to 0 h HSC (c, i), control HSC (e, j), or yHSC (e, h) levels. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.
