Fig 2: Effects of GDQ on neuronal density and caspse-3 activation within hippocampal and selected subcortical regions. Data depict the effect of GDQ on the density of neurons (NeuN positive cells, A) and apoptotic cells (activated caspase-3 positive cells, B) within the caudate nucleus, putamen, hippocampal divisions cornu ammonis (CA) 1/2, CA3, CA4 and dentate gyrus (DG), thalamic medial nucleus (MN) and medial geniculate nucleus (MGN) in sham asphyxia (n = 8), asphyxia + vehicle (n = 7) and asphyxia + GDQ (n = 7) groups 7 days after umbilical cord occlusion. Data presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Bonferroni’s multiple comparisons test: *p < 0.05 vs. sham asphyxia; #p < 0.05 vs. asphyxia + vehicle; ?p < 0.05 vs. sham asphyxia and asphyxia + vehicle.

Fig 3: Impacts of elevating neurogenic factor expression on retinal ganglion cell genesis. (A) Immunofluorescent images of attached retinal organoid cultures infected with LV-GFP or LV-ATOH7f after 4-day Dox induction at day 39. Co-labeling for viral vector markers with POU4F and NF145 at day 38, or POU4F1 and RBPMS at day 41 shows high correspondence of LV-ATOH7f infection and RGC marker expression. Scale bars, 20 µm. (B) Representative flow cytometry profiles of dissociated cells at day 39 from LV-GFP, LV-ATOH7f, and LV-NEP infected retinal organoids after 4-day Dox induction. Cells were co-labeled for RGC marker POU4F and viral marker Flag for LV-ATOH7f, or GFP for LV-GFP and LV-NEP, respectively. (C,D) Bar graphs show flow cytometry quantification of RGC markers POU4F and ISL1 among total cells (C) and viral infected cells (D) at day 39. (E) Quantification of cultured monolayer cells from LV-GFP, LV-AEP, and LV-NEP infected retinal organoids at day 47 after 7-day Dox induction. Percentages of RGC marker ISL1, DCX, and NeuN positive cells among viral infected GFP+ cells are shown. For (C–E) Mean ± S.E.M. of n = 3 independent samples. One-way ANOVA, ****p < 0.0001, ***p < 0.001, **p < 0.01. “*” is used to represent levels or ranges of probability, i.e. statistical significance.

Fig 4: Expression of C1q/TNF-related protein 9 (CTRP9) and adiponectin receptor 1 (AdipoR1) after intracerebral hemorrhage (ICH). a Representative western blot images and quantitative analyses of CTRP9 time course after ICH. b Representative western blot images and quantitative analyses of AdipoR1 time course after ICH. Values are expressed as mean ± SD. *p < 0.05, **p < 0.01 vs. sham group. N = 6. c Double immunofluorescence staining for AdipoR1 (red) in microglia (Iba-1, green), neurons (NeuN, green), and astrocytes (GFAP, green) in right basal cortex 24 h after ICH. Scale bar = 50 µm. N = 2. DAPI, 4',6-diamidino-2-phenylindole; Iba-1, ionized calcium binding adaptor molecule-1; NeuN, neuronal nuclear; GFAP, glial fibrillary acidic protein

Fig 5: Hydrogel-evoked FBRs vary and exhibit cellular features of CNS wound response.a Schematic and chemical structures of synthetic hydrogels used as tools to study CNS FBR. b Experimental model of in vivo injections into caudate putamen (CP) of mouse forebrain. c, d Survey and detail images of stromal and inflammatory cells (CD13), astrocytes (GFAP), and neurons (NeuN) at 1 week after injections of PBS, hydrogels, or L-NIO-induced stroke. e–g Quantification of total immunohistochemical staining. e Total CD13 in CP. not significant (NS), ***P < 0.0001 versus PBS injection, one-way ANOVA with Bonferroni. f Total CD13 in either non-neural or neural tissue compartments. NS or ***P < 0.0001 versus PBS injection, two-way ANOVA with Bonferroni. g Total GFAP in CP. NS or ***P < 0.0001 versus DCHMO, one-way ANOVA with Bonferroni. All graphs are mean ± s.e.m with individual data points superimposed showing n = 10, 10, 12, and 4 mice per group for PBS, DCHMO, DCHK, and L-NIO, respectively. AU arbitrary units, G hydrogel, IC internal capsule.