Alexa fluor 555

SH-SY5Y cells were seeded on slides in 12-well culture plates. After drug treatment, the cells were fixed with ice-cold methanol for 5 min, blocked with 1% bovine serum albumin (BSA) for 30 min, and then incubated with primary antibodies against LC3 (1:200) and P62 (1:100) at 4°C overnight. The next day, the slides were incubated with Alexa Fluor 488 (Bioss Antibodies, bs-0295G-AF488, Beijing, China) or Alexa Fluor 555 (Bioss Antibodies, bs-0296G-AF555, Beijing, China) secondary antibodies (1:500) at 37°C for 1 h and incubated with DAPI (Boster, AR1177, Wuhan, China) for 5 min. Fluorescence was observed with a fluorescence microscope (Olympus, BX53, Tokyo, Japan).

Cells were plated on coverslips for 24 h. Cells were fixed with 4% paraformaldehyde, incubated in 0.3% Triton X-100 for 15 min, followed by blocking with 5% goat serum. The cells were then incubated with primary antibodies against YAP at 4 °C overnight, followed by appropriate secondary antibodies (Alexa Fluor 555; Bioss). The nuclei were counterstained with DAPI (Beyotime). The immunofluorescent signals were detected by Microscope (DM6000B).