Fig. 5: GRAIL interacts with sirtuin 1 (Sirt1) and enhances its ubiquitination.
From: E3 ubiquitin ligase Grail promotes hepatic steatosis through Sirt1 inhibition
A–D Endogenous Grail interacts with endogenous Sirt1. Extracts from AML12 and HepG2 cells were prepared, immunoprecipitated with anti-Grail, anti-Sirt1, or rabbit anti-immunoglobulin G (anti-IgG) antibodies, and analyzed using anti-Grail and anti-Sirt1 antibodies. E, F Lysates from HepG2 (GRAIL overexpression or silencing) stable cell lines were harvested and subjected to immunoprecipitation (IP) with Sirt1 antibody and analyzed for ubiquitylation by immunoblotting with indicated antibody. G, H HEK293 cells were transiently transfected with HA-Ubiquitin (Ub) (wild-type (WT)), HA-Ub (K48), Flag-Sirt1, and Grail after 24 h. Lysates were harvested and subjected to IP with Flag antibody and analyzed for ubiquitylation by immunoblotting with indicated antibody.
