Yvad cmk

Human OC cells A2780 were purchased from ATCC (Manassas, VA, USA) and cultured at 37 °C with 5% CO₂ in RPMI 1640 medium (Hyclone, Logan, UT, USA) containing 10% fetal bovine serum (Hyclone). To evaluate the effect of FOSL2 on OC cells, the A2780 cells were treated with either pcDNA4.0-FOSL2 plasmid or FOSL2 shRNA. The cells were randomly allocated into five groups: control, pcDNA4.0 (OE-NC), pcDNA4.0-FOSL2 (OE-FOSL2), control shRNA (shCTRL), and FOSL2 shRNA (shFOSL2). After 48 h, the OC cell function and the expression of inflammasome-associated proteins were assessed to determine the effect of FOSL2 on the formation of inflammasome and the apoptosis of OC cells. Furthermore, to confirm the role of inflammasomes in inhibiting OC cell apoptosis, Ac-Yvad-CMK was used to block inflammasome activation. For this experiment, cells were randomly divided into four groups: control, control shRNA (shCTRL), FOSL2 shRNA (shFOSL2), and FOSL2 shRNA + Yvad-CMK (400,012, Sigma, 20 μM) (shFOSL2 + Yvad-CMK).

NPCs were treated with MCM or recombinant mouse IL-1β (rIL-1β; R&D Systems) 2 days after plating as a monolayer. Unless otherwise indicated, aMCM refers to 100% aMCM. In the indicated experiments, 50 ng/ml recombinant mouse IL-1RA (R&D Systems) was added to NPC cultures simultaneously with the switch to MCM or rIL-1β treatment. For inhibition of IL-1β cleavage in microglia, 20 μM of the caspase-1 inhibitor y-VAD-CMK (Sigma) was added to microglia at the time of activation, and conditioned media was collected after 24 h.