(A) Schematic representation of the fasting experimental design.

(B to E) Blood was drawn from healthy individuals in the fed and in the fasting state and analyzed by CyTOF. (B) Multidimensional CyTOF data were clustered using viSNE. (C) Heatmap shows mean markers expression on cell clusters significantly reduced during fasting. (D and E) Paired analysis of (D) total monocytes, CD14+CD16- monocytes, and CD14-CD16+ monocytes, and (E) neutrophils in human blood during the fed and the fasted state. Dotted lines indicate physiologic range.

(F) Food intake of individual mice during 4 h between ZT2 and ZT6, and percentage of food intake between ZT2 and ZT6 with regard to 24 h food intake.

(G) CyTOF analysis of blood cells from mice that were fed or fasted for 4 h. Heatmap shows mean marker expression of clusters significantly reduced by short-term fasting.

(H) Absolute numbers of Ly-6Chigh and Ly-6Clow monocytes in the blood of mice that were fed or fasted for the indicated time.

(I) Absolute numbers of neutrophils in the blood of mice that were fed or fasted for 20 h.

(J) Absolute numbers of Ly-6Chigh monocytes in the peritoneal cavity (PC), lung, spleen, liver and adipose tissue (AT) of mice that were fed or fasted for 20 h.

(K) Percentage of caspase-3/7+ cells among Ly-6Chigh monocytes in the blood and bone marrow (BM) of mice that were fed or fasted for 4 h.

(L) Numbers of bone marrow CXCR4+ pre-monocytes in mice that were fed or fasted for 20 h.

(M) Absolute numbers of bone marrow Ly-6Chigh monocytes in mice that were fed or fasted for the indicated time.

(F, H to M) Every dot represents one individual animal. Horizontal bar = mean. Vertical bar = SD. Student’s t test (D, E, I to L) or one-way analysis of variance (ANOVA) with Dunnett’s multiple comparison test (H, M) were performed. Statistical significance is indicated by ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. ns = not significant.