Autophagy is required for G-CSF responsiveness in vivo. (A) Total white cell (WC) and (B) neutrophil counts in the PB and (C) BM neutrophil counts after 6 days’ G-CSF or saline. (D) Representative flow cytometry plots from the PB of WT or Atg5^−/− FLC after G-CSF. Progenitors are enriched in lin^negc-Kit⁺ (LK⁺) populations. Quantification of mobilized LK⁺ after G-CSF in the PB and (E) BM. (F) CFU in the PB (CFU/mL of blood) and (G) BM (CFU/1 × 10⁴ BM cells) of Atg5^−/− or WT chimeras after 6 days of G-CSF or 1 dose of AMD3100 analyzed 1 hour after injection (n = 5-11, 2-3 experiments). (H) CFU in the PB (CFU/mL of blood) and (I) BM (CFU/1 × 10⁴ BM cells) of LysMCre:Atg7^fl/fl or controls after 6 days of G-CSF (n = 11-12, 3 experiments). (J) Representative plots of Annexin V and Sytox blue gated on LK⁺ cells from blood of WT and Atg5^−/− chimeras at D7 after G-CSF treatment. Percentage of Annexin V⁺ Sytox blue⁻ of blood LK⁺ cells (n = 6-8, 2 experiments). (K) Percentage of Annexin V⁺ Sytox blue⁻ cells within BM LK⁺ population after G-CSF treatment (n = 6-9, 3 experiments). (L) Percentage of Annexin V⁺ Sytox blue⁻ of blood neutrophil (CD11b⁺Gr1^hiCD45.2⁺) at D7 after G-CSF treatment (n = 7-12, 3 experiments). A Mann-Whitney U test was performed for statistical analyses (*P < .05, **P < .01, ***P < .001, ****P < .0001). For all experiments, data are expressed as mean ± SEM. (Each dot corresponds to an individual mouse.) D7, day 7.