Bms224hs

The supernatant was isolated from each culture condition and were kept at − 80 °C until further analysis. After thawing, the supernatants were used immediately for cytokine quantification using following kits according to the manufacturer’s instructions: Proteome Profiler Human Cytokine Array Kit for 36 cytokines (Biotechne, ARY005B). Secreted interleukins were identified using the following ELISA kits (sample dilution 1:5000): TNF-α (Invitrogen, BMS223HS), IL-6 (Invitrogen, BM213HS), IL-8 (Invitrogen, BMS204-3INST), IL-1β (Invitrogen, BMS224HS) and IL-10 (Invitrogen, BMS215HS). Quantification was analyzed performed by a microplate reader, TECAN M1000 pro (TECAN, Switzerland), software i-control™ (version: 2.0.10.0). The statistical analysis of the data was performed in GraphPad Prism (v8.0.2).

The corresponding ELISA kits were used to detect serum inflammatory cytokines including TNF-α (Invitrogen, KAC1751, USA, sensitivity of 3 pg/mL, range of 15-1500 pg/mL), IL-1β (Invitrogen, BMS224HS, sensitivity of 0.05 pg/mL, range of 0.16-10.0 pg/mL), IL-6 (Invitrogen, EH2IL6, sensitivity <1 pg/mL, range of 10.24-400 pg/mL), IL-8 (Invitrogen, KAC1301, sensitivity of 0.7 pg/mL, range of 7-750 pg/mL), IL-10 (Invitrogen, KHC0101, sensitivity <1 pg/mL, range of 7.8-500 pg/mL), and IL-17A (Invitrogen, KAC1591, sensitivity of 2 pg/mL, range of 15-1000 pg/mL). The procedures of ELISA assay were in accordance with the guideline from the manufacturer.