Rutin

The following antioxidants were used in this study: Trolox (Vector Laboratories, CB-1000-2), Zeaxanthin (Santa Cruz, sc-205544), L-Sodium Pyruvate (Nacalai Tesque, 06977-34), α-tocopherol (Nacalai Tesque, 34114-54), Rutin (Nacalai Tesque, 30319-04), N-Acetyl-L-cysteine (Nacalai Tesque, 00512-84) and Ascorbic acid (Nacalai Tesque, 03420-52). Ascorbic acid (100 mM) was prepared in water and stored at −20 °C. Zeaxanthin (2 mM), α-tocopherol (10 mM), Rutin (10 mM) and NAC (100 mM) were prepared in DMSO and stored at −20 °C. Trolox (100 mM) and Sodium Pyruvate (100 mM) were stored at 4 °C. Each antioxidant was diluted in 500 µL of the culture medium at a final concentration before live cell imaging.

HeLa and Huh7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (D6429, Sigma-Aldrich, St. Louis, MI, USA) with 10% FBS (F7524, Sigma-Aldrich, St. Louis, MI, USA) and incubated in 5% CO₂ at 37 °C. HeLa cells stably expressing ULK1-EGFP, GFP-Atg5, GFP-WIPI, tf-LC3, and GFP-TFEB were described previously [6 (link)]. For performing LC3 flux assays, 125 nM bafilomycin A1 (023-11641, Wako, Osaka, Japan) was used. For the aggrephagy experiment, cells were treated with 5 μg/mL puromycin (160-23151, Lot PTL1732, Wako, Osaka, Japan) for 4 h and then washed with new medium. Torin-1 (475991, Calbiochem, Darmstadt, Germany) was used at a concentration of 250 nM. Rutin (30319-04, Nacalai tesque, Kyoto, Japan) and was dissolved in 42% ethanol. Quercetin (10005169, Cayman, MI, USA) was first dissolved in DMSO at a concentration of 210 mM and then dissolved with 42% ethanol.

Ferulic acid (99%) was purchased from Sigma Aldrich (St. Louis, MO, USA). Ascorbic acid and quercetin (≤100%) were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Caffeic acid (≥98%), curcumin (100%), hesperidin (90%), rutin (95.0–101.5%), and triiodothyronine (T₃) were purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Gallic acid (≥98%), chlorogenic acid (≥95%), ellagic acid (≥95%), sesamin (≥95%), epigallocatechin-3-gallate (≥96%), genistein (≥98%), daidzein (≥95%), luteolin (≥98%), theaflavin (≥98%), isoquercetin (98%), and morin (100%) were purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Resveratrol (>99.0%), fisetin (>96.0%), isorhamnetin (>95.0%), and troxerutin (90%) were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Cyanidin chloride (99%) was purchased from Nagara Science Co. Ltd. (Gifu, Japan). Apigenin (≥98%) was purchased from LKT Laboratories, Inc. (St. Paul, MN, USA). Kaempferol (98%), myricetin (98%), and naringin (98%) were purchased from BDL Pharmatech Ltd. (Shanghai, China). Hyperoside (90–100%) was purchased from Toronto Research Chemicals Inc. (Toronto, ON, Canada). Neohesperidin (≥98%) was purchased from Chem-Impex International, Inc. (Wood Dale, IL, USA). All other chemicals were commercially available and had the highest possible purity.