A549 cells were fixed with 4% paraformaldehyde for 20 minutes, followed by blocking with 5% goat serum for 1 hour. The cells were then incubated with a primary antibody targeting TWSG1 (HV536014) at a concentration of 21 ug/ml overnight at 4°C. Afterward, the cells were incubated with a secondary antibody, Goat Anti-Rabbit IgG (Alexa Fluor-488) for 50 minutes at room temperature. The localization of TWSG1 was visualized (shown in green), and nuclear DNA was stained with DAPI (shown in blue).