Extended Data Figure 2. Flow cytometry using PBMCs exhibit differential fasting and refeeding cell-surface receptor expression levels.

a, Nutrient-load dependent CD45⁺ PBMC flow cytometry distribution. Schematic representation of cytometric labelling to distinguish cell type distributions at baseline, following 24-hr. fasting and refeeding (n=19 subjects). b, Cytometric plots and gating strategies to measure cell surface markers on T helper cells. c-j, Dot and line plots show cell populations of each subject (Wilcoxon two-sided, paired analysis to compare groups, n=19 subjects). c, Flow plots of activated classical monocytes from representative subject comparing 3 nutrient conditions. The plots show relative cell population frequencies of classical monocytes (CD14^highCD16⁻) and median fluorescence intensity (MFI) of activated classical monocytes (HLADR⁺ in CD14^highCD16⁻). d, Representative flow plots showing gating and quantitation of activated CD8⁺ T cells. Plots show expression of activated CD8⁺ T cells (CD38⁺HLADR⁺ in CD8⁺) showing significant increases in refed samples compared to baseline and fasting. e, Dot and line plots showing significant blunting in refed samples compared to baseline and fasting in activated DCs (HLADR⁺ in CD16⁻CD56⁻), myeloid DCs (CD11c⁺CD123⁻) and plasmacytoid cells (CD11c⁻CD123⁺). f, Representative flow plots showing gating and quantitation of regulatory T cells and plots show no difference in Treg cells (FOXP3⁺). g, Follicular helper T cell (CD4⁺CXCR5⁺) levels show no difference in three caloricload conditions. h, Plots showing no difference in activated NK cells (HLADR⁺ in CD16⁻CD56⁺, CD16⁺CD56⁺, and CD16⁺CD56⁻). i, Plots showing no difference in immature B cells (CD19⁺CD20⁻) and mature B cells (CD19⁺CD20⁺). j, Quantifying relative cell population frequencies with specific CD8⁺ T (Tc) markers. Cell populations of Tc1 (CXCR3⁺CCR6⁻) and Tc17 (CXCR3⁻CCR6⁺) cells show no change in three caloric-load conditions. The antibody information of BD lyoplate and 18-color panel and gating strategy of flow cytometry is shown in Supplementary Tables 1 and 2 and Supplementary Data3.