Figure 4. Co-localization of the mutant EPHA2-Myc proteins with the cis-golgi apparatus in MDCK cells. Ectopically expressed EPHA2-Myc
protein (cyan) in MDCK cells was detected using an anti-Myc primary antibody and Cy5-conjugated secondary antibody; the cis-golgi
apparatus (red) was detected using an anti-GM130 primary antibody against the cis-golgi marker, GM130, and Alexa Flour 555-conjugated
secondary antibody. Nuclei (blue) were labeled with DAPI. A: Cells expressing EPHA2-Myc protein with p.P584L (second row), p.A959T (third row), or p.V972GfsX39 (fourth row) mutation
show peripheral and cytoplasmic localization of the protein similar to that of the wild-type protein (first row). No co-localization
with the cis-golgi apparatus was observed in cells expressing these mutant proteins (Merge). B: Cells expressing EPHA2-Myc protein carrying a p.T940I or p.D942fsXC71 mutation show mis-localization of the protein in the
perinuclear space (first and third row) and co-localization (white) with the cis-golgi apparatus (Merge, first and third row;
arrow). A few cells expressing these mutant proteins show peripheral and cytoplasmic localization of the protein (second and
fourth row), with some co-localization with the cis-golgi apparatus (Merge, fourth row, arrow). Representative images from
two independent experiments are shown. Scale-bar=20 µm.