Fig. 4: Cdo1 deficiency exacerbates hepatocytes steatosis in vitro.

From a–g Cdo1^flox/flox loci-containing primary hepatocytes were infected with adenovirus harboring Cre (AD-Cre) for the ablation of Cdo1. After 24 h, cells were treated with a mixture of 0.6 mM FFAs (oleate and palmitate) at a final ratio of 2:1 for 24 h. Then cells were harvested and analyzed. n = 3 independent biological replicates for western blotting. n = 6 independent biological replicates for other experiments. a Nile red and Hoechst staining of the cells. Scale bars, 50 μm. b Western blotting of the cell lysates. c The mRNA levels of the indicated genes. d MitoScene and Hoechst staining of the cells. Scale bars, 50 μm. e The mitochondrial DNA (mitoDNA) levels of the cells. f Oxygen consumption rate (OCR) of the cells. FCCP, mitochondrial uncoupler. R&A, rotenone and antimycin A. g Fatty acid oxidation (FAO) levels of the cells. h–n HepG2 cells were transfected with the indicated small interfering RNA (siRNA). After 24 h, cells were treated with a mixture of FFAs as described above for 24 h. Then cells were harvested and analyzed. n = 3 independent biological replicates for western blotting. n = 6 independent biological replicates for other experiments. h Nile red and Hoechst staining of the cells. Scale bars, 50 μm. i Western blotting of the cell lysates. j The mRNA levels of the indicated genes. k MitoScene and Hoechst staining of the cells. Scale bars, 25 μm. l The mitoDNA levels in HepG2 cells. m OCR in HepG2 cells. n Fatty acid oxidation (FAO) levels in HepG2 cells. Experiments were performed 3 times and similar results were obtained in a, b, d, h, i and k. Unpaired two-tailed t tests were performed in c, e and g; one-way analysis of variance plus Tukey’s post hoc tests were performed in j, l and n; two-way analysis of variance plus Tukey’s post hoc tests were performed in f and m. All data show the means ± SD. Source data are provided as a Source Data file.