Fluorescent confocal image of Hela cell stained with NKX1-1 Antibody (Center).Hela cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with NKX1-1 primary antibody (1:25, 1 h at 37℃). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:400, 50 min at 37℃).Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7units/ml, 1 h at 37℃). Nuclei were counterstained with DAPI (blue) (10 µg/ml, 10 min). NKX1-1 immunoreactivity is localized to Nucleus significantly.