Mtt powder

Amyloid beta-peptide (1-42) (PA4391) was purchased from Ontores Biotechnologies (Zhejiang, China). Recombinant human Dectin-1 (rhDectin-1) was purchased from Sino Biological (10215-HNCH; Beijing, China). HA-tag (51064-2-AP) and Flag-tag (20543-1-AP) were ordered from Proteintech (Shanghai, China). Biotin (D8150) and Biotinylated-Aβ₄₂ (Bio-Aβ₄₂) was obtained from GL Biochem (Shanghai, China). Penicillin/Streptomycin and Opti-MEM were ordered from Gibco (Carlsbad, CA, USA). Annexin V-FITC/PI Apoptosis Detection Kit was purchased from BD Biosciences (San Diego, CA, USA). Bovine serum albumin (BSA), Dulbecco's modified Eagle's medium (DMEM) and Dimethyl sulfoxide (DMSO) were ordered from Sigma (St. Louis, MO, USA). MTT powder, reactive oxygen species assay kit (ROS), JC-1 assay kit, RIPA lysis buffer and Hoechst 33258 were bought from Beyotime Institute of Biotechnology (Shanghai, China). PVDF membrane, ECL Plus and Western Blot Marker were bought from Bio-Rad. The list of antibodies used and their source is presented in the supplementary informationTable S1. Sequence Information of β-Amyloid (1-42) used in this study was shown in the Supplementary InformationTable S2.

A 5 mg/mL MTT solution was prepared using MTT powder (Beyotime, Shanghai, China) and PBS. Cells (5000–7500 cells/well) were seeded into a 96-well plate. Following overnight incubation, varying concentrations of K784−8160 culture supernatants were added to each well, and the cells were further cultured for an additional 48 h and 72 h. Next, a 10% MTT solution was added to each well. After an incubation period of 1–3 h at 37 °C in the dark, the supernatant was discarded, and the formazan crystals were dissolved using DMSO. Absorbance was measured at 570 nm using a microplate reader (TECAN, Zurich, Switzerland).