Fig. 1

Analysis of overall immune responses of red blood cells to STIF by proteomics and flow cytometry. (A) Schematic diagram of the 6-day STIF program and expression dynamics of DEPs. Upper, Schematic illustration of the STIF program that includes fasting for 6 days and gradual refeeding for 6 days (n = 31). Bottom, Expression dynamics of all DEPs between STIF day 0, STIF day 6 and refeeding day 6. Protein abundance at the later time points was normalized to STIF day 0 as relative expression. (B) Differential protein expression and GO analysis of red blood cells. Top, Volcano plot of the red blood cell proteome. Lower panel, GO enrichment analysis of the differential proteome. Left, fasting day 6 vs. fasting day 0, right, refeeding day 6 vs. fasting day 0. The color bar indicates -log₁₀(P value). The bubble size indicates the number of DEPs annotated to a GO term. The horizontal axis is the fold enrichment (gene ratio/background gene ratio) in GO enrichment analysis. The vertical axis is the name of the GO term. (C) GSEA on the regulation of immune system processes. (D) Annotated heatmap of protein abundances related to positive regulation of immune system process (left) and negative regulation of immune system process (right). (E) Flow cytometric quantitation of the immune function of red blood cells (GPA⁺) marked by CD35. Fasting day 0 (dark blue), fasting day 6 (sky blue), refeeding day 6 (purple), n = 31/group