Figure 5. IL-1β secretion is independent of MTA.

BMDM were primed for 4 h with 10 ng/mL LPS in the absence or presence of 200 µM or 500 µM MTA, then washed and treated in the presence (MTA in Sig 2) or absence of 200 µM MTA, inflammasome inhibitors YVAD or KCl and NLRP3 agonists 3 mM ATP, 2000 U/mL SLO or 20 µM nigericin for 30 min. Supernatants were collected and analyzed by ELISA (A) or TCA-precipitated, resolved by SDS-PAGE along with cell lysates, and transferred to PVDF (B). Blots were sequentially probed with 3ZD anti-IL-1β mAb, EPR3057 anti-HMGB1 rabbit mAb and anti-actin mAb coupled with relevant HRP-conjugated secondary antibodies. The graph is the mean ± sem of 4 experiments while the blot is a representative blot from 3 independent experiments.