Glycine

Quantitative ChIP analysis was performed as previously described 20 (link). Cells were cultured to 80-90% confluency and then crosslinked with 1% formaldehyde (Sigma) at RT for 10 min. After neutralization with glycine (125 nmol/L, Sangon), cells were lysed in SDS lysis buffer. The antibody against FLAG (Sigma) was used to pull down the DNA and protein complex, and the purified DNA was subjected to qRT-PCR. We designed primers encompassing the predicted RBP-Jk binding sites in promoter regions of SLUG and GAS1. Subsequently, 1% of total chromatin supernatant before immunoprecipitation was used as input. The primers used were provided in Table S4.

HAuCl₄^.4H₂O was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Cellulase, pepsin, trypsin and AA were obtained from Yuanye Biotechnology Co., Ltd. (Shanghai, China). Histidine, threonine, lysine, glycine, glutathione (GSH), maltose, sucrose, glucose and metal ions were acquired from Sangon Biotechnology Co., Ltd. (Shanghai, China). All reagents were of analytical purity and used directly. Milli-Q purified water prepared by the PR03200 ultra-pure water meter (Zhongshan Keningte Cleaning Supplies Co., Ltd.) was utilized in all experiments.

Hydrochloric acid (HCl), tris(hydroxymethyl) aminomethane (Tris), MgCl₂, NaCl, ZnCl₂, ascorbic acid were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Exo III, guanosine monophosphate, cytidine monophosphate, glycine, and lactose were obtained from Shanghai Sangon Biotechnology Co., Ltd. (Shanghai, China). Melamine was purchased from Aladdin Reagents Co., Ltd. (Shanghai, China). All these chemicals were used without further treatment. All solutions were prepared by using ultrapure water (resistivity  18.2 MΩ cm) obtained from a Milli-Q water purification system (Millipore Corp., Bedford, MA, USA). The HPLC-purified DNA probes were obtained from Shanghai Sangon Biotechnology Co., Ltd. (Shanghai, China) and their sequences were listed as follows:
HP: 5′-GTC TGT TTT TTT TCT CAG ACT CTT TTT TTT-3′
MB-DNA: 5′-AGA AA X AA X ACA GAC-MB-3′
X denotes the AP site of spacer C3.
The MB-DNA and HP were directly used and diluted in 50 mM Tris-HCl (pH = 7.2, containing 100 mM NaCl, 5 mM MgCl₂) to give the stock solution of 10 µM.

Chloral hydrate, sodium chloride injection, normal saline, and methanol were bought from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Glycine, TRIS buffer, and sodium dodecyl sulfate (SDS) was purchased from Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). Skim milk powder was purchased from BBI Life Sciences (Shanghai, China). 30% acrylamide and ammonium persulfate (AP) were bought from Beijing Dingguo Changsheng Biotechnology Co., Ltd. (Beijing, China). Tetramethylethylenediamine (TEMED) was provided by Sigma (St. Louis, MO, United States). GeneView (Ste. Genevieve, MO, United States) provided the enhanced chemiluminescence (ECL) Kit. The Milli-Q (18.2 MΩ) ultra-pure water system (Millipore, Billerica, MA, United States) was used to prepare pure water. Mouse serum interferon gamma (IFN-γ), IL-10, IL-12p70, IL-17A, TNF-α, IL-6, monocyte chemoattractant protein-1 (MCP1), and IL-1β enzyme-linked immunosorbent assay (ELISA) kits were purchased from Bender (Gruenberg, Germany).
DF was prepared in our laboratory [Batch No.: 20180522. The DF had a rhodojaponin III content of 41.6% and a rhodojaponin VI content of 13.6%, as determined by the linear curve method (Yao et al., 2019 )]. Dexamethasone was purchased from GlpBio (Batch No.: GC40775. Montclair, CA, United States).

ATP and AMP were purchased from Aladdin (Shanghai, China) in the form of sodium salt. Magnesium acetate was from Sigma (St. Louis, MO, USA). Bromothymol blue sodium salt, glycine, and other reagents all came from Sangon Biotech Corp. (Shanghai, China). Plastic cuvettes were purchased from Centome Corp. (Chengdu, China).