Extended Data Fig. 7: Transplanted enTsOrg shows significant gene expression changes related to neural functional recovery.

a, Immunofluorescence analysis of p-PI3K, p-AKT and p-GSK3β expression in neurons of enTsOrg and sOrg 7 weeks post-transplantation. b, Violin plot of the expression levels of UCHL1, MFN2, MFN1, DMN1L, VDAC1 and OPA1 determined by ST-seq (n = 2). c, qPCR analysis of UCHL1, MFN2, VDAC1 and OPA1 genes in spinal cord tissues, expression levels were normalized to the sOrg group using the 2^–ΔΔCt method. Two-sided Welch’s t test (n = 4, UCHL1, VDAC1; n = 3, OPA1; n = 10, MFN2, biological replicates). d, Western blot results of the protein expression of MFN2, VDAC1 and Drp1 in each group 7 wpi with the bar graphs representing quantitative analysis. e, Representative immunofluorescence images of UCH-L1 expression in neurons of 7 and 14 weeks spinal cord tissues post-transplantation. The magnified areas are shown in the white boxes; white arrowheads indicate UCH-L1 + /NeuN+ cells. f, g, Immunofluorescence analysis of dynamic marker (MFN2, Dlp1 and OPA1) expression in longitudinal spinal cord slices. The magnified areas are shown in the white boxes. h, Expression and distribution of MFN2 in organoid transplant areas. i, Immunofluorescence analysis of mitochondrial marker VDAC1 expression in longitudinal spinal cord slices of enTsOrg and sOrg group 7 weeks post transplantation. j, Immunofluorescence analysis of SV2 expression in longitudinal spinal cord slices of enTsOrg and sOrg group 7 weeks post transplantation. Data represent the mean ± s.e.m. (c). Scale bars, 100 μm in (a, g, j). wpi, weeks post injury; p-PI3K, phosphor-phosphoinositide 3-kinase; AKT, protein kinase B; GSK3β, Glycogen Synthase Kinase 3; Vsx2, visual system homeobox 2; UCH-L1, ubiquitin c-terminal hydrolase L1; MFN2, mitofusin 2; Dlp1, dynamin-like protein 1; OPA1, optic atrophy 1; VDAC1, voltage dependent anion channel 1; SV2, synaptic vesicle glycoprotein.

Source data