Figure 4. FH1FH2mDia2 binds directly to... | Rockefeller University Press

Figure 4.

$Figure 4. FH1FH2mDia2 binds directly to MTs. (A) SDS gel analysis of the cosedimentation of His- (His-mDia2), GST- (GST-mDia2), or untagged (mDia2) FH1FH2mDia2 or GST alone (GST) with taxol-assembled MTs or BSA. Matching input and pellet fractions were resuspended in SDS loading buffer and run on 10% SDS-PAGE followed by Coomassie staining. tub, tubulin. (B) Concentration dependence of FH1FH2mDia2 binding to taxol stabilized MTs (1.5 μM tubulin). Error bars show SEM of three independent experiments. (C) SDS gel analysis of MT cosedimentation assay, as described in A, with His-tagged versions of wild-type (WT) and mutant (K853A, I704A, and W630A) FH1FH2mDia2. (D) Concentration dependence of wild-type and mutant His-FH1FH2mDia2 binding to MTs. Input material (I) and MT (or BSA) pellets material are shown. (E) MT cosedimentation analyses of wild-type and mutant His-FH1FH2mDia2 with intact and sheared MTs. (F) Immunostaining of His-FH1FH2mDia2 on immobilized MTs as described in Materials and methods. Bar, 10 μm.$

FH1FH2mDia2 binds directly to MTs. (A) SDS gel analysis of the cosedimentation of His- (His-mDia2), GST- (GST-mDia2), or untagged (mDia2) FH1FH2mDia2 or GST alone (GST) with taxol-assembled MTs or BSA. Matching input and pellet fractions were resuspended in SDS loading buffer and run on 10% SDS-PAGE followed by Coomassie staining. tub, tubulin. (B) Concentration dependence of FH1FH2mDia2 binding to taxol stabilized MTs (1.5 μM tubulin). Error bars show SEM of three independent experiments. (C) SDS gel analysis of MT cosedimentation assay, as described in A, with His-tagged versions of wild-type (WT) and mutant (K853A, I704A, and W630A) FH1FH2mDia2. (D) Concentration dependence of wild-type and mutant His-FH1FH2mDia2 binding to MTs. Input material (I) and MT (or BSA) pellets material are shown. (E) MT cosedimentation analyses of wild-type and mutant His-FH1FH2mDia2 with intact and sheared MTs. (F) Immunostaining of His-FH1FH2mDia2 on immobilized MTs as described in Materials and methods. Bar, 10 μm.