Extended Data Fig. 6: HRI NTD mediates its interactions with DELE1.
From: DELE1 oligomerization promotes integrated stress response activation
a. Co-immunoprecipitation of FLAG-tagged HRI and GFP tagged WT DELE1CTD (CTD), and different DELE1 mutants. Anti-GFP antibody and Anti-FLAG antibody were used to detect DELE1 and HRI, respectively. b. The AlphaFold2 model of HRI. The HRI N terminal domain (NTD: 1-160) is colored blue, and the HRI C terminal domain (CTD: 161- 630) is colored yellow. The disordered kinase insertion region (239-375) from the AlphaFold2 model was deleted and instead shown as dashed line. c. Structural alignment for the kinase domains of PKR and HRI. The crystal structure of PKR kinase domain (PDB: 2A19) is colored purple, and the HRI kinase domain is yellow. d. Schematic of DELE1 and HRI constructs used for co-immunoprecipitation (co-IP). e. Co-IP of GFP-tagged WT DELE1 C-terminal domain (CTD), and different Flag tagged HRI constructs including full-length HRI (FL), HRI N-terminal domain (NTD), and HRI C-terminal domain (CTD). An anti-GFP antibody and anti-FLAG antibody were used to detect DELE1 and HRI, respectively. Left: Immunoblotting of designated proteins from IP fractions. Right: Immunoblotting of designated proteins from whole cell lysis (WCL) fractions. In 6a and 6e, Data shown are representative of two independent experiments.
