Migfilin localizes at both focal adhesions and late endosomes/lysosomes. (A and B) KP4 cells were costained with anti-migfilin antibody and anti-paxillin antibody (A) or anti-Rab7 antibody (B), and analyzed from Z-series images taken at various heights by confocal microscopy. Extended upper planes showed that migfilin puncta were concentrated in the perinuclear regions and co-localized with Rab 7 (open arrows), but not with paxillin (solid arrows). Extended bottom planes showed that migfilin puncta were accumulated in focal adhesions and co-localized with paxillin (solid arrows), but not with Rab7 (open arrows). (C) KP4 cells expressing mCherry-Migfilin were stained with anti-paxillin antibody and analyzed from Z-series images taken at various heights by confocal microscopy as A. (D) Living KP4 cells expressing mCherry-Migfilin and GFP-Rab7 were analyzed from Z-series images taken at various heights by confocal microscopy as B.