Fig. 3: Fasting enhances GR transcriptional activity and increases expression of the transcriptional suppressor ZBTB16.

a, Volcano plot depicting genes that are differentially expressed between TMX and TMX plus fasting treated MCF7 xenografts. Differential gene expression was determined by two-sided Wald test. b, Gene set enrichment analysis for Hallmark pathways. Shown are the pathways that are differentially enriched upon treatment with TMX or TMX plus fasting. Normalized enrichment score (NES) was calculated by weighted Kolmogorov–Smirnov test and P value was determined by permutation-based testing with multiple Benjamini–Hochberg hypothesis correction. DN, downregulation; UP, upregulation. c, Enrichment plot of a pan-cancer GR-activity signature for all depicted conditions. NES was calculated by weighted Kolmogorov–Smirnov test and P value determined by permutation-based testing with multiple Benjamini–Hochberg hypothesis correction. d, Differential expression analyses for GR-activity signature genes, based on log₂-transformed fold change ranking, comparing TMX and TMX plus fasting conditions. e, Snapshots of H3K27ac, ERα, GR, PR and JUN ChIP–seq signal at the ZBTB16 gene locus in TMX-treated and TMX plus fasting-treated xenografts. f, Enrichment plot for GR-activity signature in matched tumour samples of patients with breast cancer, before and after five days of FMD. NES was calculated by weighted Kolmogorov–Smirnov test and P value was determined by permutation-based testing with multiple Benjamini–Hochberg hypothesis correction. g, Correlation plot between of G2M and E2F Hallmarks, and the GR-activity signature in matched tumour samples from patients with breast cancer, before and after five days of FMD. Two-sided Spearman’s linear correlation between gene set variation analysis (GSVA) enrichment scores of the indicated gene sets was calculated, and R and P values are shown.