Extended Data Figure 1. Initial analysis of RNA-seq data acquired from the 3 nutritional-load conditions.

a, Individual points (symbols) and means±SEM (lines) of glucose, insulin and growth hormone levels following 24-hr. fasting and 3-hrs. following a fixed caloric meal. Dot plots represent mean±SEM with value of each subject (n=26–28 subjects). The values represent average of duplicates. Two-sided, paired Student’s t-tests. Sera glucose (n=28), Sera insulin (n=26), Sera growth hormone (n=26). b, Table showing number of DE genes (p<0.05) identified in the indicated comparisons. PBMCs’ RNA from 21 subjects used to generate RNA-seq data. c, Unsupervised principal component analysis (PCA) performed on DE genes (p<0.05) for indicated comparisons (n=21 subjects). d, Combined PCA analysis of the top 1000 DE genes (p<0.05) from all 3 groups. e, Top 10 pathways in which the DE genes (p<0.05) from the indicated comparisons. The q values (p values adjusted for false discovery rate (FDR)) from the enrichment result are represented by negative log10 scale (x axis). The most significant pathways predominantly align with lymphocyte and T cell differentiation and activation comparison with refed state. f-g, Box and whisker plots show range of fold change of the subset of 114 DE genes (p<0.05, RNA-seq analysis) that were either downregulated (f) or upregulated (g) to a greater degree following the 24-hr. fast vs. refeeding than baseline (overnight fast) vs. refeeding. The box and whiskers plots show median and upper/lower quartile of the relative gene expression. The whiskers show Turkey distribution and the outlier levels are shown as individual genes (n=21 subjects). h, Pathway enrichment analysis of 844 DE genes exclusively identified in the 24-hr. fasted versus refed state shown with q values (p value adjusted for FDR) for each pathway represented by negative log 10 scale (x axis). Statistical Source Data of Extended Data Fig. 1