Immunofluorescence Analysis of DEK and β-Tubulin

MKN-1 gastric cancer cells grown on coverslips was fixed with 4% paraformaldehyde in PBS for 10 min at room temperature and permeabilized with 0.5% TritonX-100 for 10 min. Then washed again with PBS and blocking was performed with 3% Albumin Bovine V (A8020, Solarbio, Beijing, China) for one hour at the room temperature (RT). Primary antibodies against DEK (1:50; BD Biosciences, USA) and β-Tubulin (1:50; Santa Cruz Biotechnology, USA) were incubated with cells at 4°C overnight. After more washes, cells were incubated with Alexa Fluor® 488 Goat Anti-Rabbit IgG (H + C) (A11008, 1:1000, Invitrogen, USA) and Alexa Fluor® 568 Goat Anti-Mouse IgG (H + L) (A11004, 1:1000, Invitrogen, USA) for 1 h. Subsequently, cells were washing again with PBS and counterstained with 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (C1006, Beyotime, China). Coverslips were mounted with Antifade Mounting Medium (P0126, Beyotime, China) [17 (link)]. Finally, the immunofluorescence signals were visualized and recorded by Leica SP5II confocal microscope.