Fig. 6
Proteasome inhibition promotes AGR2 autophagic degradation via cargo adaptor NBR1. a Immunofluorescence analysis of AGR2 and p62 localization treated by MG132 for 24 h in A549 cells. For confocal microscopy, AGR2 and p62 were immunostained, with nuclei stained with DAPI. Scale bar = 10 μm. b The interaction between AGR2 and p62 was evaluated in MG132-treated A549 cells by immunoprecipitation with anti-AGR2 antibody and immunoblotting (IB) with anti-p62 antibody. c Knockdown of p62 by siRNA, the change of AGR2 protein in MG132-treated A549 cells was analyzed by western blot. d The interaction between AGR2 and other autophagy receptors response to MG132 was evaluated by immunoprecipitation. e Immunofluorescence analysis of AGR2 and NBR1 localization. Scale bar = 10 μm. f Knockdown of NBR1 by siRNA, the change of AGR2 protein in MG132-treated A549 cells, was analyzed by western blot. g Flag-AGR2 and His-NBR1 or His-NBR1-ΔUBA (deletion of the ubiquitin-associated domain) were co-transfected into HEK293 cells, and the lysates from transfected cells were subjected to immunoprecipitation (IP) with anti-Flag antibody followed by immunoblotting (IB) analysis with anti-His antibody. h The Pearson’s correlation test was used to analyze the link between AGR2 and NBR1 from GEO database
