Localization of uPA, cytokeratin and actin in MCF10A neoT cells. Colocalization of anti-uPA antibody and anti-CK MAb (A). MCF-10A neoT cells were stained with anti-uPA MAb (green fluorescence, Alexa-488) and anti-CK MAb (red fluorescence, Alexa-555) and imaged sequentially in a line-interlace mode to eliminate cross talk between the channels. The threshold level for this display was set arbitrarily. Pixels above the threshold in both channels (blue color, right panel) and the contour plot are shown for MCF10A neoT cell cluster (inset C) demonstrating strong co-localization in contrast to adherent MCF-10A neoT cells (B). Anti-CK Mab triggered actin reorganization in MCF-10A neoT cells. MCF-10A neoT cells, treated (C) and non-treated (D) with anti-CK MAb, were cultured on fibronectin for 2 h and stained with phalloidin. Actin bundles are seen in anti-CK8 MAb treated cells but not in non-treated cells.