As029

Methyl isonipecotate (≥70%, Sigma, USA), thiophene-2-thiol (≥99%, Sigma, USA), ethyl acetate (≥99.8%, Energy Chemical, China), croconic acid (>95%, Aladdin, China), n-butanol (≥99%, Aladdin, China), toluene (98%, Yishi Chemical, China), N-hydroxysuccinimide (NHS, 98%, Aladdin, China), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, Aladdin, China), imiquimod (R837, 99%, Meryer, China), aB2MG (ABclonal, China), 4,6-biphenyl-2-phenylindole (DAPI, Beyotime, China). For immunostaining experiments, the following antibodies were used: rabbit anti-forkhead box O4 (FOXO4) (A17978, ABclonal, China), rabbit anti-p16^ink4a (A0262, ABclonal, China), rabbit anti-p21CIP1 Rabbit pAb (A1483, ABclonal, China), rabbit anti-lamin B1 (A16909, ABclonal, China), rabbit anti-p53 (A0263, ABclonal, China), rabbit anti-decoy receptor 2 (DcR2) (A6136, ABclonal, China), mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (AC002, ABclonal, China), goat anti-rabbit IgG H&L Alexa Fluor^® 594 (ab150080, Abcam, UK), HRP goat anti-rabbit IgG (H+L) (AS029, ABclonal, China) and HRP goat anti-mouse IgG (H+L) (ab205719, Abcam, UK).

According to previously reported method [23 (link)], total proteins were isolated from cells or tissues, separated with sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane. Prior to incubated with antibodies, the membranes were cut and then incubated with anti-rabbit ATG5 antibody (1:1000, #A0203, ABclonal, China), anti-rabbit Beclin-1 antibody (1:1000, #A7353, ABclonal), anti-rabbit transforming growth factor (TGF)-β1 antibody (1:1000, #A7040, ABclonal), anti-rabbit α- smooth muscle actin (SMA) antibody (1:1000, #A17910, ABclonal) and anti-rabbit GAPDH antibody (1:1000, #AC001, ABclonal) overnight at 4 °C. After washing, the membrane was cultured with the second antibody (1:1000, AS029, ABclonal) for 60 min at 37 °C. An enhanced chemiluminescence reagent kit (#K1230, ApexBio, Shanghai, China) was used to visualize the bands.

The slices were treated with H₂O₂, boiled twice, and washed according to reported descriptions [28 (link)]. The slices were incubated overnight with rabbit antibodies, α-SMA (1:800, #A17910, ABclonal), TGF-β1 (1:800, #A7040, ABclonal) and LC3B (1:800, (#MA5-37852, 1:800, ThermoFisher Scientific, China) at 4 °C. After washing, the slices were incubated with the secondary antibody (1:1000, AS029, ABclonal) for 120 min at 37 °C, then stained with diaminobenzidine, dehydrated, cleared and sealed. All analysis of immunohistochemical staining sections was performed using ImageJ software.