Endothelial cells resident in a monolayer culture were immunofluorescently labeled with Alexa Fluor 488 conjugated to goat secondary antibodies that target mouse anti-NPCP (nuclear pore complex protein; green fluorescence). The cells were also counterstained with MitoTracker Red CMXRos (mitochondria; red fluorescence) and Alexa Fluor 350 conjugated to phalloidin (filamentous actin; blue). Images were recorded in grayscale with a Hamamatsu ORCA-AG camera system coupled to an Olympus BX-51 microscope equipped with bandpass emission fluorescence filter optical blocks provided by Omega Filters. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.