Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Dec 17;7:0551. doi: 10.34133/research.0551

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2024 Jiashu Yang et al.

Exclusive licensee Science and Technology Review Publishing House. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License (CC BY 4.0).

PMC Copyright notice

Fig. 6. — HSP90α inhibition promoted the ubiquitination of NLRP3 mediated by Cbl-b. (A) BMDMs treated with 10 μM polydatin or GA were stimulated with 100 ng/ml LPS for 3 h, or (B) HEK293T cells transfected with flag-NLRP3 were treated with 10 μM polydatin or GA. NLRP3 ubiquitination level was analyzed by immunoblot. (C) HEK293T cells were transfected with various combinations (above lanes) of plasmids encoding flag-NLRP3, GFP-HSP90α, and HA-Ub, and immunoblot analysis of NLRP3 ubiquitination (detected by anti-HA antibody) in cell lysates immunoprecipitated with anti-flag agarose. (D) HEK293T cells were transfected with various combinations (above lanes) of plasmids encoding flag-NLRP3, GFP-HSP90α and HA-Ub, HA-Ub K48 only, or K63 only. Immunoblot analysis of NLRP3 ubiquitination (detected by anti-HA antibody) in cell lysates immunoprecipitated with anti-flag agarose. (E) Schematic representation of NLRP3 WT and deletion mutants. (F) HEK293T cells were transfected with various combinations (above lanes) of plasmids encoding NLRP3 truncations as indicated. Immunoblot analysis of interaction between HSP90α and NLRP3 truncations in lysates immunoprecipitated with anti-flag agarose. (G) HEK293T cells were transfected with various combinations (above lanes) of plasmids encoding HA-Ub, GFP-HSP90α, and NLRP3 truncations as indicated. Immunoblot analysis of ubiquitination of NLRP3 truncations (detected by anti-HA antibody) in cell lysates immunoprecipitated with anti-flag agarose. (H) BMDMs treated with CQ and MG132 were added 10 μM polydatin or GA and then exposed to 100 ng/ml LPS for 3 h. An endogenous IP with NLRP3 antibody was performed. (I) BMDMs infected with HSP90α Cas9 lentivirus were treated with 10 μM polydatin or GA, followed by 100 ng/ml LPS for 3 h. NLRP3 ubiquitination level was analyzed by immunoblot. Data are representative of 3 independent experiments.