Ti e a1 si confocal microscope

Alexa Fluor 555 Phalloidin (Cat# 8953S, Cell Signaling Technology) was commercially obtained and used to stain F-actin in cells. In brief, an average of 1.5 × 10⁵ cells were seeded into a 35-mm confocal dish. After cultivation for 48 h, cells were rinsed with ice-cold PBS, fixed with 4% paraformaldehyde (Cat# AR-0211, DINGGUO CHANGSHENG BIOTECHNOLOGY C.LTD., Beijing, China) at 4°C for 30 min, permeabilized with 0.2% Triton X-100 (Cat# AR-0341, DINGGUO CHANGSHENG BIOTECHNOLOGY C.LTD.) for 10 min, and blocked with normal goat serum (Cat# 16210064, Thermo Fisher Scientific) for 1 h. Then, cells were incubated with 0.33 μM phalloidin at room temperature for 15 min, and cell nucleus were stained with DAPI (Cat# H-1200-10, Vector Laboratories, CA, USA) for 5 min. Images of stained cells were captured using a TI-E + A1 SI confocal microscope (Nikon, Tokyo, Japan).