Fig. 1
FER phosphorylation of CRMP2 impairs its microtubule bundling activity. a Paclitaxel-stabilized rhodamine-labeled microtubules were incubated in the absence or presence of CRMP2 or pCRMP2 (phosphorylated by FER kinase) at room temperature for 40 min before fluorescence microscopy was performed and images were obtained. Bar plots represent the mean + s.e.m. of microtubule width in µm from at least 150 individual microtubules per condition tested. Data presented are typical of at least three independent replicates. Scale bar is 10 µm. b Electron microscopy images of microtubules in the absence or presence of recombinant 6His-CRMP2. The dashed rectangles in the left panel are the areas zoomed in on the right panel. Scale bar is 100 nm. c Recombinant 6His-CRMP2 (residues 13–516) was purified and incubated with paclitaxel-stabilized microtubules for 30 min at room temperature. Subsequently, the samples were ultracentrifuged at 200,000×g for 30 min before analysis using SDS-PAGE and coomassie staining. S: supernatant, P: pellet. d Paclitaxel-stabilized rhodamine-labeled microtubules were incubated in the absence or presence of CRMP2 at room temperature for 40 min. CRMP2 localization was revealed by anti-CRMP2 antibody. Scale bar is 10 µm
