Mouse il 6 flex set

High glucose Dulbecco’s modified Eagle’s Medium (DMEM, GIBCO, PA, USA). Fetal bovine serum (FBS, EVERY GREEN, China). Penicillin/streptomycin (P/S, GIBCO, USA). AG490 (HY-12000, MCE). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) (Solarbio, Beijing, China). Docetaxel (114977–28-5, aladdin, shanghai, china). IL-6 (Mouse IL-6 Flex Set, 558301, BD), IL-17A (Mouse IL-17A Flex Set, 560283, BD), TNF-α (Mouse TNF Flex Set, 558299, BD), IFN-γ (Mouse IFN-γ Flex Set, 558296, BD). DAPI staining reagent (G1012, servicebio, Wuhan, China). SZYY obtained from C. officinalis Planting Base in Taiping Town, Xixia County, Nanyang City, Henan Province, China.

The measurement of nitric oxide (NO) in the supernatants was taken using the modified Griess reagent (Sigma, G4410). Samples (100 μl) were plated in a flat 96‐well plates (Corning), mixed with equal volumes of freshly prepared Griess reagent, and incubated for 15 min at room temperature. The plate was read at 540 nm using a microplate reader (LabSystems). The cytokine release to the supernatants was measured using BD™ Cytometric Bead Array (CBA) following the manufacturer’s protocol and detected using a BD FACSVerse flow cytometer. The following kits were used to detect indicated cytokines: Mouse IL‐6 Flex Set (BD, 558301), Mouse TNF Flex Set (BD, 558299), Mouse MCP‐1 Flex Set (BD, 558342), and Mouse IL‐10 Flex Set (BD, 558300). The cytokine level in the supernatant was reflected by the median fluorescence intensity (MFI) of PE of the corresponding capture beads.