Fig. 2.

Generation of a novel 3D hepatic steatosis model and increases in CYP4A and ER stress in a steatosis model. a Brief outline of the generation of a 3D hepatic steatosis model after culture with high glucose and palmitate for 5 days. b Intracellular lipids were stained with Oil red O (top) and Nile red (bottom) and quantified by Nile red fluorescence intensity (right); n = 3. c mRNA and d Protein expression levels of gluconeogenesis- and lipogenesis-related genes were determined in control or glucose/palmitate, or tunicamycin (ER stressor)-treated 3D spheroids. β-Actin was used as an internal control for mRNA and protein expression; n = 3. e The triglyceride concentration was quantified by absorbance in each group; n = 3. f Glucose uptake was quantified by fluorescence intensity in each group; n = 3. g mRNA expression levels, h Protein expression levels, and i Activity of CYP4A were determined in control, glucose/palmitate-treated, or tunicamycin-treated 3D spheroids; n = 3. j Western blot analysis of ER stress markers in each condition. β-Actin was used as an internal control. *p < 0.05, **p < 0.01, and ***p < 0.001