Super-resolution localization of polarity regulators at the plasma membrane

Super-resolution localization of polarity regulators at the plasma membrane will involve a) immunization of llama's with MDCK cells to generate a MDCK-immune library, b) production and purification of the membrane anchoring component for selection of specific nanobodies, c) selection and characterization of nanobodies binding to anchoring proteins, d) imaging of membrane anchoring of the spindle in 3D by super-resolution microscopy of 2D MDCK cell cultures, and e) quantitative characterization of the organization of the spindle orientation by anchoring complexes and the effect of EGF thereon.