Fig. 5. mTOR inactivation in SCs results in accelerated AxD.

a, Left: Scheme of Phospho Explorer antibody array analysis with array images from the experiment using pooled groups of control and axotomized nerve segments. Right: Upregulation of phosphorylation targets representing mTORC1/2 (magenta squares) and AMPK (turquoise triangles) induction in axotomized nerve segments in comparison to upregulation of c-Jun and p-ErbB2 (Tyr877) (n=6 uninjured nerve segments and n=6 injured nerve segments from n=6 mice, each symbol represents the fold change value in comparison to the pooled uninjured control nerve group), b, c, Western blot analysis (cropped blot images) of lysates from uninjured nerve segments and axotomized distal sciatic nerve stumps from C57Bl/6J mice reflecting mTORC1 (b) and mTORC2 activity (c) at different times following nerve transection. Individual lanes represent pooled data from at least three mice. d-g, Representative immunofluorescence for the indicated markers on longitudinal frozen sections from control uninjured nerves and axotomized distal sciatic nerve stumps at the shown post-injury time points. Arrows depict colocalization. Scale bars: 50μm. The experiments were reproduced three times independently with similar results. h, Quantitative analysis of relative axon survival in distal sciatic nerve stumps 36h after axotomy in mice with the indicated genotypes (Error bars represent s.e.m. n=3 mice with the genotype mTOR^fl/fl and n=4 mice with the genotype mTOR^fl/fl; P0^Cre). Statistical evaluation was performed using Student’s t-test, unpaired, two-tailed, i, Representative confocal projections of whole-mounted distal sciatic nerve stumps from mice with the indicated genotypes show increased fragmentation of transected YFP⁺ axons in the mTOR-deficient preparation. Scale bar: 100μm. The experiment was reproduced three times independently with similar results. j, Left: Representative electron micrographs of transverse sections from distal nerve stumps of mice with the indicated genotypes 48h after sciatic nerve transection with pseudocoloring of intact (turquoise) and degenerated (magenta) myelinated fibers show increased axon death in mTOR-deficient sample. Scale bar: 10μm. The experiment was reproduced three times independently with similar results. Right: Representative immunofluorescence of transverse frozen sections from 48h axotomized sciatic nerve stumps of mice with the indicated genotypes show decreased NF-H immunoreactivity in the mTOR-deficient preparation (blue: DAPI). Scale bar: 50μm. The experiment was reproduced three times independently with similar results.