Annexin 5 fitc kit

Phellinus ribis was collected in Jinan, Shandong province (China). DEAE-cellulose 23 was acquired from Serva (Heidelberg, Germany) and Superdex 30 was purchased from Amersham Biosciences (Uppsala, Sweden). Hydrogen peroxide (H₂O₂, 30%, V/V), NaNO₃, and ascorbic acid (Vc) were purchased from Guoyao (Shanghai, China). Standard monosaccharides, including d-mannose (Man), d-xylose (Xyl), d-glucose (Glc), d-galactose (Gal), larabinose (Ara), l-rhamnose (Rha), and d-glucuronic acid (GlcU), were purchased from sigma (St. Louis, MO, USA). 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and DMSO were purchased from Sigma (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were acquired from Gibco (Carlsbad, CA, USA). Aβ_25–35 was purchased from Abcam. Annexin V-FITC kit was from Bestbio (Shanghai, China) and JC-1 kit was from Kaiji (Nanjing, China). The BCA protein kit, TBST buffer, rabbit polyclonal antibodies against Cytc, rabbit polyclonal antibodies against caspase 3, and mouse monoclonal against caspase 3 were from Biyuntian (Wuhan, China); anti-GAPDH was from Solarbio (Beijing, China). All other reagents used in the experiment were of analytically pure agent grade.

Cell apoptosis assay was performed using the Annexin V‐FITC Kit (BestBio) according to the manufacturer's instructions. TUNEL assay was carried out as described in Data S1. The cell cycle analysis was performed using the cell cycle detection regent kit (BestBio) according to the manufacturer's instructions.

After being treated with STAT3 decoy ODN or scrambled ODN for 12 h, cells were harvested, washed with PBS at 4 °C, and resuspended in 100 μL binding buffer (1 × 10⁶ cells/mL) containing 5 μL of Annexin V-FITC and 10 μL of PI (propidium iodide) using an Annexin V–FITC kit (BestBio, Shanghai, China). After incubation for 10–15 min in the dark at room temperature, these cells were analyzed by flow cytometry.