Fig. 6

Structural reorganization of the axonal cytoskeleton following injury. A Representative scanning electron microscopy images of AS in control and axons subjected to injury (scale bar chamber: 200 µm, close-up: 20 µm, right horizontal: 5 µm, swellings: 2 µm). B Frequency distribution of the number of AS measured per axon (n = 5). C Frequency distribution of inter-swelling distances for control and injured axons. The distribution was fitted with three Gaussian curves with means of 3.09 µm, 6.12 µm and 13.87 µm (variances: 0.75, 3.02, 8,76 µm, respectively, control n = 57, injury n = 181). D SIM images of axons transduced with mem-mCherry and immunostained for βII-Spectrin or actin (scale bar: 2 µm). E Identification of AS and shaft ROIs, masking for surface (mem-mCherry), identification of actin or βII-Spectrin particles and measurements of the distances between the closest neighbours. Distances in shaft vs distances in swellings for actin and βII-Spectrin (n > 14). F SIM images of axons transduced with mem-mCherry and immunostained for βIII-tubulin or pNF. Images were processed and magnitude of directionality measured and plotted in polar plots (scale bar upper: 5 µm, middle: 2 µm). G Representative image of the axons immunostained for CAMSAP2 and acetylated tubulin after injury (scale bar: 20 µm, close-up: 5 µm). H Frequency distribution of the periodicity of CAMSAP2 staining along the axons after injury (n = 96). I Representative image (z-projection) of an axon transduced with mem-mCherry and immunostained for CAMSAP2 and acetylated tubulin (scale bar: 10 µm, close-up: 5 µm). Data show mean ± SEM (**p < 0.01, ***p < 0.001). Statistical comparisons were performed using the student t-test (E). See also Additional file 13: Fig. S4