Figure 1. Kank interacts and is... | Rockefeller University Press

Figure 1.

$Figure 1. Kank interacts and is partially colocalized with IRSp53 in vivo. (A) Coimmunoprecipitation of Kank and IRSp53 in VMRC-RCW cells. Approximately 600 µg of total protein lysate (shown on the right) was used for immunoprecipitation (IP) with a preimmune serum or with a polyclonal antibody against Kank or IRSp53. The proteins in the immunoprecipitates were detected by Western blotting (WB; arrowheads). (B) Immunostaining of Kank and IRSp53 in VMRC-RCW cells. Confocal laser microscopic images were obtained for Kank (green) and IRSp53 (red) using anti-Kank (monoclonal) and anti-IRSp53 (polyclonal) antibodies, respectively, and their colocalization at the cell periphery (arrowheads) is shown in yellow. (C) Immunostaining of Kank and WAVE2 in VMRC-RCW cells. Confocal laser microscopic images were obtained for Kank (green) and WAVE2 (red) using anti-Kank (polyclonal) or anti-WAVE2 (goat polyclonal) antibodies, respectively, and their colocalization at the cell periphery (arrowheads) is shown in a merged image (yellow). (D) Fractionation of HeLa cells. The proteins in the membrane and cytosolic fractions were separated, and those indicated were detected by Western blotting. EGF receptor is a marker for the membrane, and β-actin and β-tubulin are markers for the cytosol. Bars, 10 µm.$

Kank interacts and is partially colocalized with IRSp53 in vivo. (A) Coimmunoprecipitation of Kank and IRSp53 in VMRC-RCW cells. Approximately 600 µg of total protein lysate (shown on the right) was used for immunoprecipitation (IP) with a preimmune serum or with a polyclonal antibody against Kank or IRSp53. The proteins in the immunoprecipitates were detected by Western blotting (WB; arrowheads). (B) Immunostaining of Kank and IRSp53 in VMRC-RCW cells. Confocal laser microscopic images were obtained for Kank (green) and IRSp53 (red) using anti-Kank (monoclonal) and anti-IRSp53 (polyclonal) antibodies, respectively, and their colocalization at the cell periphery (arrowheads) is shown in yellow. (C) Immunostaining of Kank and WAVE2 in VMRC-RCW cells. Confocal laser microscopic images were obtained for Kank (green) and WAVE2 (red) using anti-Kank (polyclonal) or anti-WAVE2 (goat polyclonal) antibodies, respectively, and their colocalization at the cell periphery (arrowheads) is shown in a merged image (yellow). (D) Fractionation of HeLa cells. The proteins in the membrane and cytosolic fractions were separated, and those indicated were detected by Western blotting. EGF receptor is a marker for the membrane, and β-actin and β-tubulin are markers for the cytosol. Bars, 10 µm.