Journal: bioRxiv

Article Title: Rab11FIP2 controls NLRP3 inflammasome activation through Rab11b

doi: 10.1101/2025.05.19.654879

Figure Lengend Snippet: NLRP3 binds to FIP2 via its KMKK motif ( A ) Immunoblot of Flag-NLRP3 pulldown in lysates from HEK293T cells co-transfected with Flag-NLRP3, EGFP-FIP2, and ECFP-Rab11. ( B ) Immunoblot of Flag-FIP2 pulldowns in lysates from HEK293T cells co-transfected with EGFP-NLRP3, ECFP-Rab11, and HA-ASC. ( C ) Immunoblot of Flag-NLRP3, Flag-NLRP3 132-1036 and Flag-NLRP3 152-1036 pulldowns in lysates form HEK293T cells co-transfected with EGFP-FIP2 ( D ) Immunoblot of Flag-NLRP3 or Flag-NLRP3 AAAA pulldowns from HEK293T cells co-transfected with EGFP-FIP2. ( E ) Immunoblot of Flag-NLRP3 pulldowns from HEK293T cells co-transfected with EGFP-FIP2 or EGFP-FIP2 I481E . ( F ) Immunoblot of Flag-FIP2, Flag-FIP2 1-192 or Flag-FIP2 193-512 pulldowns from HEK293T cells co-transfected with EGFP-NLRP3. HEK293T cells were transiently transfected with expression plasmids encoding the indicated fusion proteins for 48 h, or 24 (D – F) before immunoprecipitation by anti-Flag affinity agarose. The Empty-Flag-vector were used to ensure equal plasmid DNA amounts.

Article Snippet: Generation of a stable THP-1 cell line co-expressing Flag-FIP2, Flag-Rab11a or Flag-Rab11b THP-1 expressing lentiviral encoding FIP2 was made by cloning FIP2 into the bicistronic lentiviral expression vector pLVX-EF1α-IRES-Puro-Vector (Takara, 631988) and co-transfected with packaging plasmids psPAX2 and pMD2.G, kindly provided by the TronoLab (Addgene plasmids 12260 and 12259, to produce pseudo viral particles in HEK293T cells (ATTC CRL-3216™).

Techniques: Western Blot, Transfection, Expressing, Immunoprecipitation, Plasmid Preparation