Figure 3. Microtubule-binding mutations... | Rockefeller University Press

Figure 3.

$Figure 3. Microtubule-binding mutations on Moesin do not perturb FERM–CERMAD interaction or Moesin’s association with the plasma membrane and cortical actin network. (A) The GST fusion protein bound to magnetic glutathione beads was allowed to interact with the FERM domain in solution. Bead-bound fraction (P, pellet) was separated from the unbound fraction (S, supernatant) using a magnet. Both FERM wild type and FERM K212,213M associate with CERMAD in a 1:1 stoichiometric ratio. (B) FERM wild type and FERM K212,213M domain of Moesin fused to GFP were expressed in S2 cells. Bar, 10 µm. (C) Cells expressing the indicated Moesin constructs were plated on concanavalin A–treated glass coverslips to allow firm spreading. GFP expression and F-actin staining with Texas red–phalloidin are shown. Bars, 5 µm. (D) Quantification of rounding in cells transfected with the indicated constructs. Boxes show top and bottom quartiles, horizontal lines show median values, and vertical lines show minimal and maximal values. Moe, Moesin; WT, wild type.$

Microtubule-binding mutations on Moesin do not perturb FERM–CERMAD interaction or Moesin’s association with the plasma membrane and cortical actin network. (A) The GST fusion protein bound to magnetic glutathione beads was allowed to interact with the FERM domain in solution. Bead-bound fraction (P, pellet) was separated from the unbound fraction (S, supernatant) using a magnet. Both FERM wild type and FERM K_212,213M associate with CERMAD in a 1:1 stoichiometric ratio. (B) FERM wild type and FERM K_212,213M domain of Moesin fused to GFP were expressed in S2 cells. Bar, 10 µm. (C) Cells expressing the indicated Moesin constructs were plated on concanavalin A–treated glass coverslips to allow firm spreading. GFP expression and F-actin staining with Texas red–phalloidin are shown. Bars, 5 µm. (D) Quantification of rounding in cells transfected with the indicated constructs. Boxes show top and bottom quartiles, horizontal lines show median values, and vertical lines show minimal and maximal values. Moe, Moesin; WT, wild type.