Fig. 5: RAS GTPases co-localize with AFDN.
From: Afadin couples RAS GTPases to the polarity rheostat Scribble
a Expression of wild-type KRAS does not alter AFDN or SCRIB localization. EGFP-KRAS was expressed in MCF7 cells, and localization of endogenous AFDN and SCRIB was detected by immunostaining and confocal microscopy. Projections of z-stacks are at right and top. Position of associated z projections are shown with dashed yellow lines in the merged image. Scale bars represent 10 µm. b KRAS-G12V co-localizes with AFDN and SCRIB in MCF7 cells. Following expression of EGFP-KRAS-G12V, endogenous AFDN and SCRIB were detected by immunostaining. Cells expressing KRAS are found in the monolayer (1), detaching from the monolayer (2) or on top of the monolayer (3). Projections of z-stacks are at right and bottom of the merged images and their position is marked with a dashed yellow line. Scale bars represent 10 µm. c Activated RAS GTPases recruit AFDN to the cell membrane. Cherry-AFDN was co-expressed with EGFP-tagged RAS or RAP GTPases in HeLa cells. EGFP alone or EGFP-tagged wild-type KRAS with AFDN were controls, as was Cherry alone with activated KRAS-G12V. Scale bars represent 10 µm.
