Fig. 6: ENDOG activates ER stress by translocating to the ER and binding with Bip.

a ENDOG binding proteins detected by IP-MASS. Protein Acce.: Protein accession. EmPAI: exponentially modified protein abundance index. b Co-IP analyses. HepG2 cells were transfected with ENDOG-Flag plasmid for 48 h. L: long exposure; S: short exposure. c–e Representative images of costaining of ENDOG with Grp75/Calnexin/Bip after 200 μM oleic acid treatment for 24 h. f, g Representative images of in situ proximity ligation assay (PLA) and quantitative results of PLA puncta per cell. HepG2 Cells were treated with 200 μM oleic acid for 24 h. n = 12 biologically independent samples. h Subcellular fraction isolation by density gradient centrifugation. HepG2 cells were treated with 200 μM oleic acid for 24 h. ER, endoplasmic reticulum fraction; Mito, mitochondrial fraction; p, precursor ENDOG; m, mature ENDOG. i The ER location of ENDOG is analyzed by ER enrichment Kit. HepG2 cells were treated with 200 μM oleic acid for 24 h. Total, total cell lysis; ER, endoplasmic reticulum enriched fraction. j Co-IP analyses. HepG2 cells were cotransfected with Bip-Myc and pK-Myc/ENDOG plasmids for 48 h. k Endogenous Co-IP analyses in wild-type and ENDOG knockout HepG2 cells. l Endogenous Co-IP analyses. HepG2 cells were treated with 200 μM oleic acid for 24 h. Three experiments were repeated independently with similar results in (b, c, d, e, h, i, j, k, l). Statistical significance was determined by unpaired Student’s t-test (two-tailed) in (g); error bars are mean ± SD. Source data and exact P value were provided in a Source data file. ***P < 0.001.