Immunofluorescence staining of PCFT. HeLa cells were transiently transfected with cDNA of wild-type and PCFT mutants and labeled with a rabbit antibody targeted to human PCFT. (A) Wild-type PCFT. (B) P425R. (C) R376W. (D) S318R. Green fluorescence indicates localization of PCFT, while the blue fluorescence (panels A-B) or red fluorescence (panels C,D) shows nucleus counterstained by DAPI or PI, respectively. The image is a representative view from at least 3 experiments. The same antibody was used for immunohistochemical and Western blot (Figure 4B) analyses. The image was obtained on Olympus 1X70 inverted Epifluorescence Microscope (Olympus) with a 60×/1.40 oil objective, recorded on a Sensicam QE cooled CCD Camera (Applied Scientific Instrumentation), acquired with software IP Lab 4.0 (BD Biosciences) and further processed with Photoshop CS (Adobe Systems).