Fluorescent image of U251 cells stained with HSPD1 antibody. U251 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated HSPD1 primary antibody (1:100, 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). Pictures were taken on a Biorevo microscope (BZ-900, Keyence). Note the highly specific localization of the HSPD1 mainly to the mitochondria, supported by Human Protein Atlas Data (http://www.proteinatlas.org/ENSG00000144381).