Scribble knockdown does not alter MCF10A cell morphology or apical-basal polarity in MCF10A acini. (a) Western blot of MCF10A cells transduced with pRetroSUPER retroviral shRNA vectors. Shown below is quantitation of Scribble knockdown relative to tubulin protein level as measured using densitometry averaged from three independent experiments. (b) DIC images of MCF10A polyclonal cell lines. (c) DIC images of MCF10A cell lines cultured in Matrigel for 10 days to form 3D acinar structures. (d) MCF10A grown in Matrigel for 20 days were fixed and processed for immunofluorescence. Columns, left to right, show representative examples from MCF10A parental, shEGFP and shScrib6 knockdown acini. Each row was stained with antibodies indicated in the left-hand panel. White arrows indicate the Golgi apparatus in Scribble knockdown cells, which appeared less condensed than in control acini.