Fig. 7: Contusion injury-induced dysregulation of intra- and extracellular mitochondria.

Differentially expressed genes (DEGs) associated with a, mitochondria complex genes; and b, mitochondria ribosomal proteins were detected (abs(Log2(Fold Change))> 0.585 and q < 0.01) in human 3D in vitro triculture model composed of induced neural stem cells derived neurons, primary astrocytes, and iPSC-derived microglia from two healthy donors YZ1 and ND41866*F (n = 6) and summarized in the over-representation analysis of c, mitochondria pathways. d, f Representative images, and e–g quantifications of Tuj1 neuronal and TOMM20 mitochondria networks in naïve neurons treated with mitochondria isolated from astrocytes activated with mitochondria isolated from YZ1 microglia that had been treated with conditioned media collected from 24 h injured neurons. h, Representative redox ratio maps of mitochondria isolated from microglia activated by conditioned media from injured neurons. Quantification of i, redox ratio, j, bound fraction, and k, a long lifetime in live mitochondria isolated from iPSC microglia, astrocytes, or neurons following N-M-A-Nt experimental design (Fig. 5a). Data presented in (a–c) were analyzed from mRNA sequencing data collected from n = 6 for sham groups and n = 6 scaffolds for injury groups from two independent experiments. Data presented in (e, g, i–k) mean ± SEM of n = 6–10 scaffolds per condition. *, **, *** indicates a significant difference (p < 0.05, 0.01, 0.001 respectively; two-way ANOVA (analysis of variance) or one-way ANOVA between control and experimental groups) (with Tukey’s post hoc test). Experiments were replicated at least three times, unless otherwise specified; metabolic images were replicated two times. Scale bar: 50 μm in d, f; 10 μm in h.