Fig. 4

Histopathological analysis of the cervical spinal cord in SOD1^G93Amice following hNSC ICV transplantation. a) Representative 3D reconstructions showing the analysed cervical SC of hNSC- vs. HBSS-treated mice (n ≥ 4/group; EXP 5). b - e) Analysis of (b) gray matter, (c) white matter, and (d) total spinal cord volume as well as e) gray/white matter ratio in the cervical tract of the SC of hNSC- vs. HBSS-treated mice at 40 dpt (n ≥ 4/group; EXP 5); parametric unpaired t-test with Welch’s correction GFAP, green; DAPI, blue. f) Astrogliosis evaluation of the cervical tract of the SC of hNSC- and HBSS-treated mice at 40 dpt (n ≥ 4/group; EXP 5); nonparametric unpaired t-test with Mann-Whitney’s correction| GFAP, green; DAPI, blue. g) Neuropathological analysis of the cervical tract of the SC assessed measuring microglial distribution in mice transplanted with hNSCs vs. HBSS at 40 dpt (n ≥ 4/group; EXP 5); parametric unpaired t-test with Welch’s correction| IBA1, green; DAPI, blue. h) Total amount of microglial cells (number of IBA1⁺ cells per mm²) in the ventral horn gray matter of the cervical SC of hNSCs- vs. HBSS-transplanted mice at 40 dpt (n ≥ 4/group; EXP 5); parametric unpaired t-test with Welch’s correction| IBA1, green; DAPI, blue. i) In vivo morphological analysis of IBA1⁺ cells at 40 dpt assessed by total number of endpoints/cell; parametric unpaired t-test with Welch’s correction (n ≥ 4/group; EXP 5). j) In vivo quantification of microglial branching at 40 dpt (n ≥ 4/group; EXP 5); nonparametric unpaired t-test with Mann-Whitney’s correction (p-value: *≤0.05). k) Representative confocal images of microglial cells within the ventral horn of the spinal cord of hNSC- vs. HBSS-treated SOD1^G93A mice.| IBA1, green; DAPI, blue. l) Phenotypic analysis and representative confocal image of microglial cells classified based on their ramified, activated (both bushy and rod-like cells), and ameboid morphology; parametric unpaired t-test with Welch’s correction (ramified and ameboid groups) and nonparametric unpaired t-test with Mann-Whitney’s correction (activated group) (n = 3/group; EXP 5; p-value: *≤0.05)| IBA1, green; DAPI, blue. m) TREM2 protein levels in the cervical tract of the SC of hNSC- and HBSS-treated animals (n ≥ 4/group; EXP 5); parametric unpaired t-test with Welch’s correction. n) MN stereological count in the cervical SC of hNSC- vs. HBSS-treated mice at 40 dpt (n ≥ 4/group; EXP 5); parametric unpaired t-test with Welch’s correction

All data are expressed as mean values ± SEM