E el h0103c

A549 and H/R-A549 cells (5x10³/well) were inoculated into 96-well plates and treated with different MAF concentrations (5, 10 and 20 µΜ) for 24 h at 37˚C. Subsequently, the cell culture medium was centrifuged at 2,000 x g for 4 min at 4˚C to remove debris and the supernatant was collected for assay. The levels of IL-6, IL-1β and IL-10 in cells were quantified using their corresponding ELISA kits (cat. no. E-EL-H0102c, E-EL-H0149c and E-EL-H0103c; Elabscience Biotechnology Co., Ltd.) according to the manufacturer's protocols. The absorbance at 450 nm was measured using a microplate reader (Thermo Fisher Scientific, Inc.).

The cells were plated in a 6-well plate and processed according to groupings. After 48 hours, the cell culture supernatant was collected, centrifuged at 10,000 g at 4 ℃ for 5 minutes, then the supernatant was obtained. Detection was performed according to the operation of the ELISA detection kit (IL-6, E-EL-H6156, IL-10, E-EL-H0103c, IFN-γ, E-EL-H0108c, Elabscience Biotechnology Co., Ltd.)

Both IL-6 and IL-10 were detected by ELISA, and the detection kits were purchased from Wuhan Elabscience Biotechnology Co., Ltd., with the item numbers of E-EL-H0102c and E-EL-H0103c, respectively. Refer to the kit instructions for specific detection steps.