Figure 2.

HSCs dynamically transition between autophagic flux states.

A Schematic for competitive transplantation of RFP⁺EGFP⁻ high flux, RFP⁺EGFP⁺ low flux, and RFP⁻EGFP⁻ no flux HSCs into irradiated recipients.

B-E Donor hematopoietic (B), B (C), T (D), and myeloid (E) cell engraftment in peripheral blood of mice receiving 25 RFP⁺EGFP⁻ high flux, RFP⁺EGFP⁺ low flux, or RFP⁻EGFP⁻ no flux HSCs with 3×10⁵ bone marrow cells (n=10–13 recipients/HSC subset in 3 experiments).

F-I HSC engraftment (F) and autophagic activity (G-I) in the bone marrow of mice from transplants in B-E 16 weeks post-transplantation (n=8–9 recipients/HSC subset in 3 experiments).

J Heat map comparing differentially expressed transcripts between RFP⁺EGFP⁻, RFP⁺EGFP⁺, and RFP⁻EGFP⁻ HSCs (≥1.5-fold change, Padj<0.05, n=3).

K Normalized enrichment scores from GSEA of most upregulated and downregulated pathways in autophagic versus non-autophagic HSCs.

L Frequency of Ki67⁺ RFP⁻EGFP⁻ or RFP⁺ CD48⁻LSK HSC/MPPs (n=3).

Data show individual mice (F-I, L) and/or mean (B-I, L) ± SEM (B-E) or SD (F-I, L). Data were assessed by a one-way repeated measures (B-E) or ordinary (F-I) ANOVA followed by Tukey’s test, or student’s t-test (L). *P≤ 0.05, **P≤ 0.01, ***P≤ 0.001, ****P≤ 0.0001.