Eclipse 80i

Relative cell numbers were assessed using a premixed water-soluble tetrazolium salt (WST-1) cell viability test (Takara, Shiga, Japan) according to the manufacturer’s instructions. The cells were seeded at a density of 1 × 10⁴ cells per well. WST-1 was added to each well, and the absorbance of the microplate at 450 nm was measured after an additional 4 h incubation. The data represent three independent experiments (n = 3). DNA-synthesizing cells were visualized using an Ethynyl deoxyuridine (EdU) kit (Invitrogen, CA, USA) following the manufacturer’s instructions. Then, the cells were washed with phosphate-buffered saline, mounted with a 4’, 6-diamidino-2-phenylindole (DAPI)-containing mounting solution (Vectashield, Vector Laboratories, Burlingame, CA, USA), and imaged by microscopy (Nikon Eclipse 80i, Tokyo, Japan). The percentage of EdU-positive cells was examined in HCC cell lines using ImageJ (Bethesda, MD, USA) software. The data represent three independent experiments (n = 3).

Cell proliferation was assessed using a premixed water-soluble tetrazolium salt (WST-1) cell viability test (Takara, Shiga, Japan) according to the manufacturer's instructions. The cells were seeded at a density of 5 × 10³ cells per well and treated with JQ1 for different durations (0 h, 24 h, 48 h, and 72 h). WST-1 was added to each well. After an additional 4 h incubation, absorbances were measured at 450 nm. The data represent three independent experiments (n = 3).
Ethynyldeoxyuridine (EdU) analysis was performed using an EdU Cell Proliferation Assay kit (Invitrogen, CA, USA), following the manufacturer’s instructions. After that, the cells were washed with phosphate-buffered saline, mounted with a 4’,6-diamidino-2-phenylindole (DAPI)-containing mounting solution (Vectashield, Vector Laboratories, Burlingame, CA, USA), and imaged by microscopy (Nikon Eclipse 80i, Tokyo, Japan). The percentage of EdU-positive cells was assessed using ImageJ (Bethesda, MD, USA) software. The data represent three independent experiments (n = 3).