Fig 2: NPC grafted into striatal stroke generate astroglia that reduce fibrotic scar and bridge lesions.a Schematic summarizing experimental approach for evaluating NPC grafting in mouse striatal stroke. b Survey IIHC images of stroke lesions at 2 weeks comparing effect of hydrogel (DCHMO) alone or grafted HA-positive NPC on stroke lesion phenotype. NPC grafted lesions show Gfap-positive lesions with reduced Cd13-positive tissue in non-neural lesion core. c Quantification of Gfap and Cd13 intensity at stroke lesions as a function of the radial distanced from the center of the lesion with mean±s.e.m. (s.e.m represented as shaded region). d, e Survey IHC images of stroke lesions at 4 weeks (d) and 8 weeks (e) comparing the effect of grafted HA-positive NPC on stroke lesion phenotype. f Quantification of total Cd13 at stroke lesion cores at 2 weeks after L-NIO injection for stroke only, hydrogel (DCHMO) alone, and NPC grafting by performing area under the curve (AUC) calculations of Cd13 traces in c. (N = 4 for L-NIO only, N = 5 for gel, N = 6 NPC graft). ***p value < 0.0005. One-way ANOVA with Tukey multiple comparison test. g Quantification of total Gfap at stroke lesions at 2 weeks after L-NIO injection for stroke only, hydrogel (DCHMO) alone, and NPC grafting by performing AUC calculations of Gfap traces in c. (N = 4 for L-NIO only, N = 5 for gel, N = 6 NPC graft) Using Gfap-HA colocalization (see Supplementary Fig. 11) the total Gfap contribution from the NPC graft and host astrocytes in the NPC graft group was determined. h Quantification of total Cd13 at stroke lesion cores at 2, 4, and 8 weeks after L-NIO injection for stroke only and NPC grafting. (N = 4, 5, 6 for L-NIO only at 2, 4, and 8 weeks, N = 6, 4, 5 for NPC grafts at 2, 4, and 8 weeks). ***p value < 0.0001, **p value < 0.005, *p value < 0.05. Two-way ANOVA with Tukey multiple comparison test. All graphs are mean ± s.e.m. AU arbitrary units.

Fig 3: (A) GFP, CFP, and RFP expression in lungs of GFAPcre-Confetti transgenic mice. (B–D) GFAP immunostaining and TDT expression in lungs of ChATcre-TDT transgenic mice. Representative image of coimmunostaining cholinergic, parasympathetic multiaxon bundle. (E–H) Representative image of a multiaxon bundle stained for GFP, TH, and MBP in lungs of GFAPcre-GFP mice. (I–L) Enlargement of box in (H). TH immunostained sympathetic nerve fibers are ensheathed by GFP expressing cells in a bundle that also contains MBP immunostained structures that do not colocalize with either GFP or TH. Scale bars, 20 µm.

Fig 4: Effect of different treatments on hippocampal GFAP immuno-histochemical levels in reactive astrocytes expressed as the mean area percentage of expression. (A) Normal group, (B) Negative control group, (C) Positive control group and (D) Test group. 400×. The bar chart (E) shows the mean ± SD of the % area of immuno-histochemical levels expression of GFAP, where Gp 1 represents the normal group, Gp 2, Gp 3 and Gp 4 represent the negative control group (received 0.9% saline), positive control group (received resveratrol suspension) and test group (received resveratrol bilosomes), respectively, where (*), (**) and (***) indicate statistically significantly different values from the control group at (p < 0.05, p < 0.01 and p < 0.001), respectively.

Fig 5: Mitochondrial transfer from neuronal cells into astrocytes. (A) Schematic showing that HA-GFP were co-cultured with SK-mitodsred at a ratio of 1:1. (B) Time-lapse live images showing mitochondria from SK-mitodsred dynamically transferring into HA-GFP. (C) Confocal images showing mitochondria from SK-mitodsred in HA-GFP after co-cultured for 24 h. (D–F) Immunostaining of TOM20 (D), ATP5A1 (E), and LAMP1 (F) after co-cultured for 24 h. (G) Flow cytometry analyzing mitochondrial transfer efficiency from SK into HA after co-cultured for 24 h. (H) H1 ESC-derived astrocytes (Ast) or neurons (hN) were labeled with GFAP::GFP or SYN1::mitoBFP. (I) Confocal images showing that mitochondria from hN-mitoBFP were observed in Ast-GFP after co-cultured for 48 h. (J) Flow cytometry analyzing mitochondrial transfer efficiency from hN cells into Ast after co-cultured for 48 h.