Fig. 5
From: Alternative splicing regulates stochastic NLRP3 activity
NLRP3 splicing is regulated on a single cell level. a Single PI-negative GM-CSF derived hMDMs were FACS-sorted into individual wells and lysed. RNA was reverse transcribed and NLRP3 full-length, NLRP3 ∆ exon 5 and HPRT encoding mRNAs were pre-amplified. Transcripts were detected with nested TaqMan assays. 187 to 192 individual cells per donor. b Quantification of the single cell NLRP3 splice pattern. Shown as the mean of three donors from a. c Scheme of the burst-kinetic of gene-expressions on single-cell level, resulting in oscillations of produced mRNA levels per gene. d Human monocyte-derived macrophages were analyzed for ASC speck formation by fluorescence microscopy after NLRP3 activation with nigericin and NLRC4 activation with bacterial product PrgI (both 1.5 h). Cell nuclei were counterstained with Draq5. Scale bar represents 50 μm. Five images per well were captured, plotted are means and SD of two replicate wells, representative of four individual experiments. Overlaid symbols represent single measurements. e IL-1β ELISpot assay of hMDMs after NLRP3 or NLRC4 inflammasome activation. Shown are two independent donors. Mean and SD of technical duplicates, two independent donors. Overlaid symbols represent single measurements. See also Supplementary Fig. 5. Source data are provided as a Source Data file
