FIGURE 4.

Analysis of the GPSM3/NLRP3 interaction using BiFC. A, HEK293 cells were transfected with plasmids expressing YN-GPSM3 (aa 1–158 of YFP fused to GPSM3) and NLRP3-YC (NLRP3 fused to aa 159–238 of YFP) and the fluorescence of reconstituted YFP fluorophore was measured at 24 h post-transfection. Cells were harvested and fluorescence quantified on a plate reader. Total DNA used for transfection was normalized using empty pcDNA3.1 vector DNA. Data are expressed as relative fluorescence units (RFU) using the means ± S.E. of at least three different experiments; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 by one-way ANOVA. B, epifluorescence microscopy images of HEK293 cells transfected with plasmids expressing the two YFP protein fusions YN-GPSM3 and NLRP3-YC (top panel) or the GFP10-NLRP3 alone (bottom panel).