Fig. 5
Nogo-B promotes lipophagy in HCC cells. a Immunofluorescence of LC3 and LD in LO2 cells treated with oxLDL for 24 h followed by starvation. b Representative immunofluorescence images (top) and quantification of co-localization areas (bottom) of RAB7 and LD, and LC3 and LD in control or Nogo-B-knockdown SK-Hep1 cells treated with oxLDL for 24 h followed by starvation. c Immunofluorescence of control and Nogo-B-overexpressing LO2 cells transfected with mCherry-EGFP-LC3 plasmid for 48 h followed by starvation. d Western blot analysis of Nogo-B, p62 and LC3 expression in starved control or Nogo-B-overexpressing LO2 cells treated with or without wortmannin (Wort) or chloroquine (CQ). e, f Co-immunoprecipitation of Nogo-B and ATG5 in e control or Nogo-B -overexpressing LO2 cells and f MHCC97H cells treated with oxLDL for 24 h followed by starvation. g Representative immunofluorescence images (left) and quantification of co-localization areas (right) of Nogo-B and ATG5 in starved control or Nogo-B-overexpressing LO2 cells. h Co-immunoprecipitation of Nogo-B and ATG5 in wile-type Nogo-B (WT) or ER-motif-deficient Nogo-B (d38) ectopically expressed LO2 cells treated with oxLDL for 24 h followed by starvation. i Representative LipidTOX staining (left) and quantification of positively stained areas (right) of control or Nogo-B-overexpressing LO2 cells transfected with ATG5 shRNAs for 48 h and treated with oxLDL for 24 h followed by starvation. j Representative image (left) and quantification of colonies (right) of control and stable Nogo-B-overexpressing LO2 cells transfected with ATG5 shRNAs for 2 weeks. (Data are presented as mean ± SEM of three independent experiments in b, g, and i–j. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file)
