Fluorescent confocal image of HeLa cells stained Phospho-KLF4-S245 antibody. HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min), then incubated Phospho-KLF4-S245 primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (5.25 mu M, 25 min). Note the highly specific localization of the phosphorylated KLF4 immunoreactivity to the nuclei but not the nucleoli or the cytoplasm.