Journal: bioRxiv

Article Title: Methionine Deprivation-induced Reprogramming of Hepatic Rhythms Is Mediated by Glucocorticoid Receptor

doi: 10.64898/2026.05.04.722637

Figure Lengend Snippet: A. Immunoblot of global H3 methylation levels in livers from mice fed on control and MD diet for three weeks. B. Immunoblot of H3 methylation levels in HepG2 cultured in medium with indicated methionine concentration for 24 hr. C. The Venn diagram depicts rhythmic H3K36me3 peaks in mice liver on control diet and MD diet measured by CUT&Tag. Rhythmic peaks were defined by comparing enrichment signals between ZT9 and ZT21 as described in Methods. D. Metagene profiles depicting H3K36me3 enrichment for 453 peaks with a stronger signal at ZT21 compared to ZT9 in control-diet fed mice (peak center ± 10 kb). E. IGV browser tracks showing H3K36me3 enrichment at Elovl3 and Elovl5 genes. Diet and Zeitgeber time for animal collection are indicated to the left, gene model is indicated below the data. F. TMM normalized expression values are presented in the graph for the Elovl5 gene. G. TMM normalized expression values are presented in the graph for the Elovl3 gene. H. The Venn diagram depicts rhythmic H3K4me3 peaks in mice liver on control diet and MD diet. Rhythmic peaks were defined by comparing enrichment signals between ZT9 and ZT21 as described in Methods. I. IGV browser tracks showing H3K4me3 and H3K36me3 enrichment at Insig2 gene. Diet and Zeitgeber time for animal collection are indicated to the left, gene model is indicated below the data. J. TMM normalized expression values are presented in the graph for the Insig2 gene. Diet group and Zeitgeber time are indicated on the graphs. See also Figure S4

Article Snippet: Human liver cancer cell HepG2 was obtained from ATCC and maintained in RPMI-1640 (GIBCO, 11875119) supplemented with 10% FBS at 37 °C with 5% CO 2 .

Techniques: Western Blot, Methylation, Control, Cell Culture, Concentration Assay, Expressing

Journal: bioRxiv

Article Title: Methionine Deprivation-induced Reprogramming of Hepatic Rhythms Is Mediated by Glucocorticoid Receptor

doi: 10.64898/2026.05.04.722637

Figure Lengend Snippet: A. Serum levels of corticosterone of six-month old male mice after three weeks on the indicated diet. The column graphs indicate mean value for 3-5 animals at indicated zeitgeber time points. Significance (**p<0.01, t test) is indicated with asterisks. B. The table shows the number of oscillating GR in liver from mice on indicated diet for three weeks. C. The Venn diagram depicts the rhythmic GR binding peaks in mice liver on control diet and MD diet detected by CUT&RUN. The rhythmic peaks were defined by comparing the enrichment signals between ZT9 and ZT21 as described in Methods. D. IGV browser tracks showing the GR enrichment at Insig2 gene. Diet and Zeitgeber time for animal collection are indicated to the left. E. Motif analysis for 3,967 GR binding peaks with a stronger signal at ZT9 compared to ZT21 in control-diet fed mice. F. Motif analysis for 2,556 GR binding peaks with a stronger signal at ZT9 compared to ZT21 in MD diet fed mice. G. On the left, the western blot depicts the protein levels of STAT5 and GR in whole cell lysis of liver. GAPDH is included as the loading control. On the right, the western blot depicts the phosphorylation levels of STAT5, protein levels of STAT5 and GR in the nuclei extraction from liver. Lamin A/C is included as the loading control. H. TMM normalized expression values are presented in the graph for the Egfr gene. Diet group and Zeitgeber time are indicated on the graphs. I. The western blot depicts the protein levels of EGFR in liver from mice after three weeks on the indicated diet. GAPDH is included as the loading control. J. The western blot depicts the protein levels of EGFR and STAT5 in whole cell lysis and nuclei extraction of HepG2 cultured in medium with high methionine concentration (100 µM) or low methionine concentration (10 µM) for 40 hours following treated with EGF (0, 100 and 200 ng/mL) for 30 min. GAPDH and Lamin A/C are included as loading control. See also Figure S5

Article Snippet: Human liver cancer cell HepG2 was obtained from ATCC and maintained in RPMI-1640 (GIBCO, 11875119) supplemented with 10% FBS at 37 °C with 5% CO 2 .

Techniques: Binding Assay, Control, Western Blot, Lysis, Phospho-proteomics, Extraction, Expressing, Cell Culture, Concentration Assay