Triton x 100

The TUNEL assay was detected with the In Situ Cell Death Detection Kit (Roche, Switzerland). The tumor tissues paraffin-embedded sections were deparaffinized in xylene, then added in a graded series of ethanol (Absolute, 95%, 75%, 5 min) and ruptured the cell membranes with 0.1%TritonX-100 (Sinopharm, China) for 8 min. Followed, the slides were rinsed in TBS three times and incubated with Equilibration Buffer for half an hour at room temperature. After that, the sections were incubated with Recombinant TdT Enzyme (Thermo Fisher, USA) in dark for 1 hour at 37°C. After incubation, the slides were washed with TBS 3 times (5 min), then incubated with DAPI at room temperature for 10 minutes, installed with antifade solution (Solarbio, S2100, Beijing), and stored at 4°C in the dark. Images were photographed at fluorescence microscope (Leica, Germany).

A549 cells were inoculated into a 96-well plate at the rate of 100 μL per well (containing 5 × 10³ cells), and the peripheral holes were filled with phosphate buffer solution. After 24 h of culturing, A549 cells were treated with TGF-β1 (5 ng/mL) and DTC (23.89 μM). The experimental cells exposed to drugs for 0 h, 24 h, and 48 h were stained, respectively. The cell culture medium in the 96-well plate was sucked out, washed twice with PBS, and 100 μL 4% paraformaldehyde (Solabio, Beijing, China) was put into each well to fix the cells at room temperature for 15 min. The stationary solution was sucked out before it was rinsed with PBS 3 times, then we put 100 μL of 0.1% Tritonx-100 (Sinopharm, Shanghai, China) into each hole, let it stand at room temperature for 10 min to increase the permeability, and rinsed it again with PBS 3 times. We then added 150 μL TRITC-phalloidin (Solabio, Beijing, China) working solution to each well, incubated it at room temperature for 30 min, and rinsed with PBS twice. Each well was stained with 150 μL Hoechest 33342 (Solabio, Beijing, China) for 5 min, washed with PBS twice, and observed under a fluorescence microscope.

Xinli No. 7 (XL7) specimens were purchased from a supermarket in Weixian, Hebei Province when they reached the maturity of the respective commercial harvesting period and immediately stored at 4 °C after sampling for subsequent processing analysis. The fruits with diseases, mechanical damage, and immaturity were removed. Hydrochloric acid, sulfuric acid, methanol, anhydrous ethanol, ethyl acetate, zinc acetate, sodium carbonate, sodium bicarbonate, sodium nitrite, aluminium nitrate, sodium chloride, sodium hydroxide (analytically pure), sucrose, potassium ferricyanide, phenolphthalein, polyethylene glycol (PEG 6000, analytically pure), acetic acid–sodium acetate buffer solution (0.1 mol/L, pH = 5.5), Triton X–100, catechol, and potassium persulfate were purchased from Sinopharm Chemical Reagent Co., Ltd. (Beijing, China). Folinol (1 mol/L), gallic acid, rutin, 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH), and 2,2′-diazo-di-3-ethylbenzothiazolin-6-sulfonic acid (ABTS) were bought from Soleibao Biotechnology Co., Ltd. (Shanghai, China).