Fluorescent confocal image of U251 cell stained with NFATC4 Antibody. U251 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with NFATC4 primary antibody (1:25, 1 h at 37°C). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:400, 50 min at 37°C). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (7units/ml, 1 h at 37°C). Nuclei were counterstained with DAPI (blue) (10 ug/ml, 10 min). NFATC4 immunoreactivity is localized to Cytoplasm significantly.