Fig 2: Lysosomal expansion but not astrocytosis and ganglioside accumulation are present in the developing postnatal primary somatosensory cortex.(A) Representative confocal images from unaffected (Sgsh+/h*) and hypomorphic MPS IIIA (Sgshh/h) mouse primary somatosensory cortex stained with LAMP-1 (green). Left panels show low magnification images, scale bar 100 µm. Right panels show high magnification of layer II/III (boxed region), scale bar 50 µm. Insets in lower right corner show single cells. Nuclei were stained with DAPI (blue). (B) Quantification of LAMP-1 stained vesicle area from high magnification images. (C) Representative confocal images stained with a marker for reactive cortical astrocytes, GFAP (red) and ganglioside GM3 (green), as described in (A). Arrows in low magnification images point to cells with intracellular staining of GM3, denoted by asterisks in high magnification images. P, pial basement membrane; WM, white matter; BV, blood vessel. (D) Quantification of GFAP immunoreactivity in the parenchyma of the somatosensory cortex. (E) Quantification of GM3 immunoreactivity in the parenchyma of the somatosensory cortex. N = 3 animals per genotype per age, 2 sections per animal, 2 regions of interest per section were quantified. All brains were collected, stained, and analyzed at the same time. Findings were confirmed on a separate cohort of N = 3 animals per genotype per age. Mean ± SD.

Fig 3: Apoptosis induced by Pb/M affects neurons and oligodendrocytes. Composites A-C show confocal images of NeuN, MBP, GFAP, AC3 and DAPI stained sections. There is colocalization of NeuN with AC3 and MBP with AC3 indicating neuronal and oligodendroglia apoptosis respectively. In C, GFAP staining (green) does not colocalize with AC3 (red) indicating that astroglia are not affected by the proapoptotic effect of Pb/M.

Fig 4: Number of ?H2AX + cells was significantly increased in temporal pole and visual cortex of demented but not in resilient when compared to control brains (a). Representative photomicrographs of ?H2AX + cells on sections from control, resilient and demented cases containing visual cortex (b). Representative immunofluorescence images of colocalization of ?H2AX + cells (brown) with neurons (red) (7c 1–3, showing a progressively zoomed-in brain region), GFAP + astrocytes (green) (c 4–6, showing GFAP alone, 7c4, ?H2AX alone, 7c5, both GFAP-?H2AX, 7c6), and IBA1 + microglia (green) (7c 7–9, showing IBA1 alone, 7c7, ?H2AX alone, 7c8, both IBA1-?H2AX, 7c9). C Control (Braak 0-II); R Resilient (Braak III/IV); D Demented (Braak III/IV); **p < 0.01; ***p < 0.001. Scale bars 1 mm, 50 µm, and 20 µm. White arrows indicate colocalization between ?H2AX+ cells and neurons, astrocytes, and microglia, respectively

Fig 5: Localization of sEH in rat brain and its expression after ischemia. Representative images show double immunofluorescent staining of sEH (red), GFAP (A, green), NeuN (B, green), and Iba1 (C, green) in cerebral cortex under normal conditions. Scale bar = 50 µm for low power images. Area in white square is shown in inset at higher power. (D,E) Western blot of sEH expression on 1 and 2 days after reperfusion in the area around the ischemic lesion. The sEH and ß-actin bands were stripped and cropped from the same gel and were developed with the same exposure. Images of the full gel are shown in Supplementary Figure S1. Comparisons between sham and MCAO groups were made by t-test (means ± SD; n = 5 rats per group; *p < 0.05, **p < 0.01 vs sham group).