Co-localization assay
According to previous publications (Jiang et al., 2019), HepG2 cells (ATCC, Virginia, USA) were cultured in DMEM containing 10% FBS. Cells were treated with DMSO or 10 μM CITCO (Sigma-Aldrich, C6240, St. Louis, MO) for 48 h. Following incubation, cells were fixed in 4% paraformaldehyde for 30 min and 0.5% triton X-100 for 10 min at room temperature. Then, cells incubated with rabbit polyclonal anti-CAR (Abcam, ab62590) and mouse monoclonal anti-YAP (R&D Systems, MAB8094, MN, USA) overnight at 4°C. Cells were stained with secondary antibodies including anti-mouse IgG Alexa Fluor 488 (Cell Signaling Technology, #4408, MA, USA) and anti-rabbit IgG Alexa Fluor 647 (Cell Signaling Technology, #4414, MA, USA). Images were acquired using a confocal microscope (Olympus FV3000, Japan).