Fig. 1: ENDOG promotes autophagic flux in hepatocytes.

a, b Western blots (a) and quantitative results (b) of autophagy-related proteins in HepG2 cells transiently transfected with ENDOG or pk-Myc (empty vector) for 48 h (n = 3 biologically independent samples, data are presented as mean values ± SD, *p < 0.05; **p < 0.01). c, d Representative images (c) and quantitative results (d) of GFP-LC3 puncta in ENDOG- or pk-Myc-transfected HepG2 cells (48 h after transfection; scale bar = 10 μm; n = 75–100 independent cells examined over three independent experiments; data are presented as mean values ± SD, ***p < 0.001). e–g. Detection and quantification of autophagic flux with the mCherry-GFP-LC3 reporter in wild-type and ENDOG-KO cell lines under indicated treatments (Starvation for 6 h; rapamycin: 1 μM for 6 h, CQ: 50 μM for 6 h). Yellow puncta, autophagosomes (mCherry⁺/GFP⁺); red puncta, autolysosomes (mCherry⁺/GFP⁻); (scale bar = 10 μm; n = 100 independent cells examined over three independent experiments; data are presented as mean values ± SD; **p < 0.01, ***p < 0.001). h, i Representative electron microscopic images (h) and quantitative results (i) of autophagic vesicles in wild-type or ENDOG knockout cells after treated with 100 nM BafA1 for 6 h (red arrow: autophagic vesicle, AL: autolysosomes, AP: autophagosomes; n = 10 independent cells; data are presented as mean values ± SD; **p < 0.01). j–m Western blots and quantitative results of LC3B, SQSTM1 and ENDOG in wild-type cells (WT), ENDOG knockout (KO) cells upon starvation (j, k) and BafA1 (l, m) treatments (n = 4 biologically independent samples) (scale bar = 10 μm; data are presented as mean values ± SD, **p < 0.01; ***p < 0.001). Source data are provided as a Source data file.