Isolation and Analysis of Hepatocytes

We isolated hepatocytes from yellow catfish liver tissues according to our recent publications [12 (link)]. After 48 h incubation, intracellular Zn and LD were detected based on the protocols of our previous studies [12 (link),31 (link)]. For detection of intracellular Zn²⁺ concentration, cells were incubated with 1 μM Newport Green DCF (Invitrogen, Carlsbad, CA, USA) for 30 min. For intracellular LD staining, cells were incubated with 5 mg/mL Bodipy (Invitrogen, Carlsbad, CA, USA) for 30 min. Fluorescence was imaged using laser scanning confocal microscopy (Leica, Wetzlar, Germany). We used the commercial assay kits (Nanjing Jian Cheng Bioengineering Institute, Nanjing, China) to analyze intracellular TG content. The mRNA levels of genes involved in lipid metabolism were examined by quantitative real-time PCR (Q-PCR). Primers are given in the Supplementary Table S6.