Fig 2: Aromatase (Aro) expression in astrocytes in glia-enriched subcultures. Cells were isolated from P0–P5 newborn rats. Aro (red) is strongly expressed in GFAP-positive astrocytes (green) and colocalizes with the nuclear staining (DAPI, blue). Fibrous astrocytes with small somata and long, thin unbranched processes (a–h) show strong nuclear Aro expression (a,c,d, white arrowheads). Cytoplasmic Aro expression is weaker, and the long thin processes do not show Aro immunopositivity. Protoplasmic astrocytes (i–p) with bigger somata and short, thick and frequently branched processes also display nuclear Aro expression (i,k,l, white arrowheads). Weak immunopositivity is detectable in the processes. The nuclear Aro signal distributes evenly or in a dotted pattern (c,k, white arrowheads) or strongly concentrates to certain parts of the nucleus (g) in both astrocyte subtypes. (White arrowheads indicate a selected representative cell and its nucleus with the Aro signal). Scale bar: 100 µm.

Fig 3: Histological analysis performed on paraffin-embedded slices of resected GB of a rat.A. H&E staining confirmed the presence of GB characterized by central tumor necrosis (1), tumor (2) and the peritumoral zone with infiltrating cancer cells (3) surrounded by healthy brain tissue (4). B. Immunohistochemistry for Ki67 indicated that GB is highly proliferative, except for the necrotic tumor core: tumor necrosis (1), tumor (2), peritumoral zone with infiltrating cancer cells (3) and healthy brain tissue (4). C-D. Immunohistochemistry for GFAP and Cx43 demonstrated GFAP-positive reactive astrocytes and enhanced Cx43 expression at the peritumoral zone.

Fig 4: Immunolocalization of various markers in the APP/BIN1/COPS5 3xTg-AD mouse brain. Photomicrographs of different brain regions from wild-type (n = 4) and APP/BIN1/COPS5 3xTg-AD (n = 4) mice showing histopathological lesions identified via immunohistochemistry. ((A–D), (A’–D’)) Transverse sections of right half-brain (A,A’), dentate gyrus (B,B’), hippocampus (C,C’), and entorhinal cortex brain levels of wild-type mice (A–C) and APP/BIN1/COPS5 3xTg-AD mice (A’–C’), showing strong differences in expression density. (A,A’) Hemi-brain sections showing a basal level of the inflammatory marker CD11b in wild-type mice, contrasting with a highly-differentiated agglomeration pattern of immunoreactivity in the APP/BIN1/COPS5 3xTg-AD mouse brain (white arrows). (B,B’) High magnification of the dentate gyrus showing a marked reduction of GFAP-immunoreactive cells in the APP/BIN1/COPS5 3xTg-AD mouse brain, forming reactive clusters ((B’); white arrows) that are absent in wild-type control sections (B). (C,C’) A high density of ß-amyloid plaques (arrows in (C’)) is found in the APP/BIN1/COPS5 3xTg-AD mouse brain compared to controls (C), demonstrating severe neurodegeneration in transgenic mice. (D,D’) High magnification of the entorhinal cortex showing stronger CD11b immunoreactivity in the APP/BIN1/COPS5 3xTg-AD mouse brain than in controls (D’). (E) Quantification of data indicating the average percentage of immunoreactive markers in the experimental groups. * p < 0.05. Scale bars, 100 µm. Amyg, amygdala; ßA, ß-amyloid; CA1, hippocampal formation; CD11b, inflammatory marker; Ctx, cortex; DAPI, nuclear marker; DG, dentate gyrus; GFAP, glial fibrillary acidic protein; Th, thalamus; Ent Ctx, entorhinal cortex; Hypoth, hypothalamus; SN, substantia nigra; VTA, ventral tegmental area.

Fig 5: Representative HE and immunostaining images of immunodeficient RCS rat retinas implanted with iPSC-RPE monolayer assessed at 11 months post-implantation. Presence of fibrosis, immunoreactivity, and epithelial–mesenchymal transition (EMT) was assessed. (a) Retina with no surviving iPSC-RPE showing signs of inflammation and peri-membrane fibrosis indicated by white asterisk. (b) Absence of TRA-I-85 staining. (c) Retinas showing RPE65 expressing iPSC-RPE cells (white arrows) labelled for GFAP (glial cells), (d) CD68 (macrophages/microglia), (e) expression of classical mesenchymal markers a smooth muscle actin a SMA and vimentin. (f) Images in which iPSC-RPE monolayer appears to be present below the parylene membrane are either due to orientation difference in the implant placement or due to the survival of the RPE on the lower side of the parylene membrane.