Bovine albumin bsa

Coronal sections were immersed in TUNEL solution (F. Hoffmann-La Roche, Ltd., Basel, Switzerland) at 37°C for 1 h. After being blocked with bovine albumin (BSA; Sangon Biotech Co., Ltd.,
Shanghai, China) solution, sections were incubated with an anti-rabbit NeuN antibody (1:100, Proteintech Group, Inc., Wuhan, China) at 4°C overnight and then with a FITC-labeled goat
anti-rabbit secondary antibody (1:200, Proteintech Group, Inc.) for 1 h at room temperature. Afterwards, the samples were counterstained with 4’,6-diamidino-2-phenylindole (DAPI; Aladdin
Holdings Group Co., Ltd.). Finally, the tissues were sealed with anti-fluorescence quenching reagent (Solarbio Life Sciences), and images were acquired using a fluorescence microscope (400×
magnification, Olympus).

Zinc acetate and 2-methylimidazole (2-MeIM) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Glucose oxidase (GOx), ethanol, and 4,4-azodianiline were purchased from Aladdin Co., Ltd. (Shanghai, China). Dimethyl sulfoxide (DMSO), and mesitylene (trimethyl benzene, TMB) were purchased from Sigma–Aldrich (Shanghai, China) Trading Co., Ltd. Phosphate-buffered saline (PBS, 10×), and Dulbecco's modified Eagle's medium (DMEM) were purchased from KeyGEN (Nanjing, China). Fetal bovine serum (FBS) was purchased from ExCell Bio (Taicang, China). Penicillin/streptomycin (P/S, 100 ×), Bovine albumin (BSA), and DAPI stain solution were purchased from Sangon Biotech (Shanghai, China) Co., Ltd. Cell Counting Kit-8 (CCK-8) was purchased from TargetMol (Boston, MA, USA). JC-1 kit, propidium iodide (PI), and calcein-AM dyes, Annexin V-FITC/PI Apoptosis Detection Kit (Cat No. 40302ES50) were acquired from Yeasen Biotechnology (Shanghai, China). 2 × Phanta Max Master Mix, ChamQ SYBR® qPCR Master Mix (High ROX Premixed), and FastPure Cell/Tissue DNA Isolation Mini Kit were purchased from Vazyme Biotech Co., Ltd. (Nanjing, China). 0.05% trypsin solution was purchased from YIFEIXUE BIOTECH (Nanjing, China). RIPA buffer and Phenylmethylsulfonyl fluoride (PMSF) were purchased from Solarbio (Beijing, China).

Lung tissues were fixed in 4% paraformaldehyde, embedded in paraffin and sliced (5 µm). The slices were dewaxed, hydrated, and stained with hematoxylin (Solarbio Science
& Technology, Co., Ltd., Beijing, China) and eosin (Sangon Biotech Co., Ltd., Shanghai, China) (H&E) or periodic acid-Schiff (PAS; Leagene Biotechnology). Images reflecting
inflammatory or mucus secretion were obtained with a microscope BX53 (OLYMPUS). The inflammatory index was measured according to the following standards: 0 =normal; 1 =very mild; 2 =mild; 3
=moderate; 4 =marked; 5 =severe inflammation. The ratio of PAS-positive area to the total area was represented the mucus distribution (%).
For immunofluorescence analysis, slices were treated in antigen retrieval and then were blocked with 1% bovine albumin (BSA; Sangon Biotech (Shanghai) Co., Ltd., Shanghai, China). Slices
were incubated with rabbit anti-NF-κB p65 antibody (ABclonal Biotechnology Co., Ltd., Wuhan, China) and then incubated with Cy3-labeled goat anti-rabbit IgG secondary antibody (Applied
Biosystems, Inc., Foster City, CA, USA). After counterstaining with 4’,6-diamidino-2-phenylindole (Aladdin regents Co., Ltd.), the images were observed under the microscope BX53.