FIGURE 1. RNAseq Analysis Shows Distinct Separation of the 3 Caloric-load Conditions.

a, Schematic of the protocol showing intervals between fixed caloric meals and temporal research blood draws. The clinical protocol was established to perform immune cell profiling in 28 healthy human subjects. b, Volcano plots of all the genes in the indicated comparison is shown with DE genes. (p value threshold <0.001, colored as blue dots (Source Data: Statistical Source Data Fig. 1)). RNA sequencing was performed on PBMCs from 21 subjects following an overnight (baseline) and following a 24-hr. fast and 3 hrs. after refeeding. c, Venn diagram shows the number of overlapping versus distinct DE genes from baseline and 24-hr. fasting to refed comparisons. d, Top 10 pathways (q value<0.05, which depict p values adjusted for false discovery rates) from the pathway enrichment analysis of the 846 DE genes overlapping in the two comparisons (see Fig. 1C.). e, T cell specific differential pathway enrichment maps by overlaying the results of 24-hr. fasting vs. refed (green nodes) on top of the baseline vs. refed (red nodes) comparison. Each node represents distinct T cell processes or a pathway, and a single-colored node represents a pathway that was exclusively enriched by DE genes from one of the two comparisons. If common genes are annotated within two biological processes, then an edge connecting the two nodes is shown. The red and the teal-colored nodes represent pathways exclusively regulated by baseline or fasting respectively compared to refeeding. The bicolored nodes are regulated by both baseline and fasting vs. refeeding.