FIGURE 2.
PTPD1 interacts and colocalizes with KIF16B through its FERM domain. A, lysates from HEK293 cells transiently transfected with FLAG-PTPD1 were immunoprecipitated (IP) with anti-KIF16B antibody or control IgG and immunoblotted (IB) with anti-FLAG and anti-KIF16B antibodies. B, lysates from HEK293 cells transiently co-transfected with FLAG-PTPD1 or FLAG-PTPD1Δ1–325 and HA-KIF16B or HA-L1248A/F1249A mutant were immunoprecipitated with anti-HA and immunoblotted with anti-FLAG antibody. As a control, we used a lysate from untransfected cells. C and D, cells were transiently transfected with vector encoding Myc-tagged KIF16B, either wild type (C) or L1248A/F1249A mutant (D). Where indicated, a GFP vector was included in the transfection mixture (D). Twenty-four h after transfection, cells were serum-deprived overnight and then stimulated with EGF (100 ng/ml) for 30 min. Cells were subjected to double immunostaining with anti-Myc and anti-PTPD1 antibodies. Images were collected by confocal microscopy. A merged composite and magnified images are shown.