Extended Data Fig. 10. Additional behavioral, electrophysiological, and structural analysis of ACR-treated mice.

a, Behavioral analysis of C57Bl/6J mice over the course of ACR intoxication (or control treatment) for 14 days. Note accentuated deterioration of motor performance (rotarod, hanging wire, grip strength) relative to sensory performance (tail flick, hot and cold plate) (Error bars represent s.e.m. n=8 mice per group and time point, except n=7 mice for control treatment 0 days tail flick, *P=0.0024, **P<0.0001 for rotarod, *P<0.0001 for hanging wire, *P=0.0049, **P=0.0130, ***P<0.0001 for grip strength). b, Weight analysis of C57Bl/6J mice over the course of ACR intoxication (or control treatment) for 14 days (Error bars represent s.e.m. n=5 female and n=3 male control mice, n=4 female and male ACR-treated mice, *P=0.0299, **P=0.0346). c, Analysis of CMAP amplitudes recorded in gastrocnemius muscles evoked after sciatic nerve stimulation of C57Bl/6J mice following 14d of ACR treatment, or control treatment (Error bars represent s.e.m. n=8 mice per group). d, Representative electron micrographs of transverse tibial nerve sections from C57Bl/6J mice following 14d of ACR admininistration (or control treatment), with pseudocoloring of intact (turquoise) and degenerated (magenta) myelinated fiber profiles. Degenerated profiles appeared as collapsed myelinated axons with little axoplasm or axons with segregation of the axoplasm by segments of the myelin sheath (d1), deranged fibers with axoplasm constriction due to myelin infoldings and convolution (d2), and myelinated fibers with accumulation of membrane like material, multivesicular structures, and dense bodies in the axoplasm (d3). Scale bars: 2μm. e, Densities of degenerated axon profiles in tibial nerves from C57Bl/6J mice following control treatment or 14d of ACR admininistration (Error bars represent s.e.m. n=8 mice per group). f, Representative electron micrographs of transverse tibial nerve sections from C57Bl/6J mice following control treatment or 14d of ACR admininistration show normal ultrastructure of unmyelinated axons in Remak bundles (‘N’ depicts nuclei of SCs forming Remak bundles). Scale bar: 2μm. The experiment was reproduced three times independently with similar results. g, Weight analysis of ACR-treated control TSC2^fl/fl and mutant TSC2^fl/fl; iSox10^Cre mice (Error bars represent s.e.m. n=4 female and n=7 male mice (except n=6 male mice for 14 days treatment time point) with genotype TSC2^fl/fl, n=8 female and n=6 male mice with genotype TSC2^fl/fl; iSox10^Cre).

Statistical evaluation in a, b, and g was performed using multiple Student’s t-test, unpaired, two-tailed, and in c and e using Student’s t-test, unpaired, two-tailed.