Superoxide dismutase sod activity assay kit

RLs with a purity of over 90.0% were obtained from Huzhou Zijin Biotechnology Co., Ltd. (Huzhou, China) and stored at 25 °C. The RLs contained a small amount of water and soybean oil, which were residual from the fermentation process. The reactive oxygen species assay kit, the catalase (CAT) activity assay kit and the superoxide dismutase (SOD) activity assay kit were purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). Glucose and Rhodamine 123 were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Propidium iodide (PI) was purchased from Anhui Kule Biological Engineering Co., Ltd. (Hefei, China).

0.1 g of fresh leaves were taken to determine the content or enzyme activity of each substance in the plants using Proline (PRO) Content Assay Kit (Nanjing Jiancheng Institute of Biological Engineering, A107-1-1), Malondialdehyde (MDA) Assay Kit (Nanjing Jiancheng Institute of Biological Engineering, A003-3-1) and Superoxide Dismutase (SOD) Activity Assay Kit (Beijing Solarbio Science & Technology Co., Ltd., BC0170), respectively. Three biological replicates were available for each sample. The DAB staining method called diaminobenzidine method was used to detect the active site of peroxidase in cells. Three leaves each of pYL156 and pYL156: GhVIM28 were taken after NaCl stress and placed in DAB solution, darkened for 12 h, and observed after decolorization with 95% ethanol. The dark brown polymerization products represent the reaction of DAB with hydrogen peroxide. For trypan blue staining, the staining solution was first prepared according to the propor-tions of 10 mL lactic acid, 10 mL glycerol, 10 g phenol, 10 mg trypan blue (Solarbio, Beijing, T8070) and 10 mL distilled water, then soaked plant leaves in this staining solution, bath in a boiling water for 2 min, cooled at room temperature, and decolorized in chloral hydrate (1.25 g / mL). Daily replacement of decolorization solution was performed until the background color is eliminated.

Manganese chloride tetrahydrate and ammonium ceric nitrate were obtained from Aladdin (Shanghai, China). Polyvinylpyrrolidone (PVP) and N, N-dimethylformamide (DMF, ≥ 99.5%) were obtained from Macklin (Shanghai, China). 2′,7′-Dichlorofluorescin diacetate (DCFH-DA) was provided by Beyotime (Shanghai, China). Dulbecco’s modified eagle medium (DMEM), penicillin, streptomycin, trypsin and cell counting kit-8 (CCK-8) were purchased from Meilunbio (Dalian, China). Foetal bovine serum (FBS) was obtained from Gibco (Guangzhou, China). Anti-CD86, anti-CD206, anti-CD3, anti-CD4, F4/80, and Foxp3 were obtained from BioLegend (San Diego, CA, USA). Lipopolysaccharide (LPS) and Interferon-γ (IFN-γ) were purchased from Sigma-Aldrich (Santa Barbara, CA, USA). The hematoxylin and eosin (HE) staining kit, catalase (CAT) activity assay kit, and superoxide dismutase (SOD) activity assay kit were purchased from Solarbio (Beijing, China). All chemicals and reagents were used as received without additional purification. All solutions were prepared with deionised (DI) water, which was purified through an 18-MΩ system (Millipore, USA).