Additional file 2 of Quantification of NLRP3–caspase-1 complex assembly reveals a non-linear association with 30-day mortality in sepsis: a proof-of-concept pilot study

Additional file 2: In-vitro validation of the NLRP3–Caspase-1 proximity ligation assay. Experimental validation of the NLRP3–Caspase-1 PLA using THP-1-derived macrophages stimulated with canonical NLRP3 activators. Inflammasome activation was induced by LPS priming followed by ATP or Nigericin stimulation. PLA signal intensity was compared with established readouts of inflammasome activation, including caspase-1 activity, IL-1β and IL-18 release, and protein cleavage of caspase-1 and gasdermin D, demonstrating concordant activation patterns across all assays.