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).