Fig. 3

SUMO E3 ligase MAPL interacts and sumoylates NLRP3. a, b Reciprocal binding study with NLRP3 and MAPL in HEK293T. Expression and co-precipitation was determined by western blot with the indicated antibodies. c Sumoylation of FLAG-NLRP3 in the presence of the indicated proteins. FLAG-NLRP3 was purified under denaturing conditions and the presence of SUMO modification on NLRP3 was determined by immunoblot analysis of the eluate. Representative immunoblots of three independent experiments are shown. d The endogenous proximity between NLRP3 and MAPL is lost upon NLRP3 activation. Immortalized BMDMs were left untreated, primed with LPS for 4 h or treated with LPS (4 h) followed by 30 min with nigericin. PLA was performed in biological triplicate using α-NLRP3 and α-MAPL antibodies. Representative confocal microscopy images are shown for each condition. Scale bar, 10 µm