To monitor oligomerization or rearrangement of NLRP3 via intermolecular BRET, pairs of the acceptor (YFP) and donor (Renillaluciferase)-tagged NLRP3 variants were transfected into the HEK293 cell line and the BRET signal was followed in the resting state or after treatment with nigericin or buffer...
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). Functional Effects of the GPSM3/NLRP3 Interaction on NLRP3-dependent IL-1β Secretion To test ...
f, Immunoblot analysis of CASP1 cleavage in wild-type BMDMs and Ddx3xfl/flLysMcre BMDMs that constitutively express DDX3X–YFP from a cytomegalovirus promoter, treated with LPS and nigericin. Representative blots (n = 2 biologically independent experiments). Source data Extended Data Fig. 6 DDX...
The luciferase (donor)/YFP (acceptor)-tagged NLRP3 was expressed in HEK293T cells treated with control or RACK1 siRNA (Figure 6C). The BRET signals (“close” conformation) were comparable at the resting state for both siRNA-treated cells; a characteristic drop of the BRET signal was ...