Fig. 3

The LRR domain of human NLRP3 is subject to alternative splicing a Scheme of the NLRP3 exons and domains. Arrows indicate primers used in b and Supplementary Fig. 3a, b. b PCR of the NLRP3 LRR on cDNA isolated from LPS primed mouse BMDMs, pig and human PBMCs, respectively. Representative of at least three (mouse, human) or two (pig) individuals. c Immunoblot of human NLRP3 from primary human monocyte-derived macrophages (hMDM) or THP-1 cells. Either whole cell lysates or NLRP3 immunoprecipitates, using mAb targeted against the NACHT domain of NLRP3, were immunostained with a mAb targeted against the PYD to ensure NLRP3 specificity. Representative of two experiments. d Scores for the probability to function as splice acceptor and donor sites were calculated for all human NLRP3 LRR exon boundaries using SplicePort. e Number of exonic splice enhancer (ESE) sites within the exons of the LRR as predicted by RESCUE-ESE. f Scheme of a splice-switching oligo (SSO), blocking the spliceosomal access to an intron-exon boundary inducing AS. g Changes in the NLPR3 alternative splicing pattern of M0 hMDMs were induced with an exon 5 SSO. NLRP3 isoform expression analysis by qPCR. Mean and SEM of three donors (untreated: n = 2). h Cytokine secretion of morpholino-treated cells after priming with LPS (TNF) and nigericin-induced activation of the NLRP3 inflammasome (IL-1β). Mean and SEM of 3 donors (LPS only: n = 2). Each donor is plotted using a unique symbol shape. See also Supplementary Fig. 3. Source data are provided as a Source Data file