Fig 2: S1PR3 mediates nNOS/NO and oxidative stress during cerebral ischemia and reperfusion.

Fig 3: nNOS fluorescence image and NO content expression. (A,B) DAPI (blue)/nNOS (green) Representative immunofluorescence images and statistical results of mouse brain tissue slices after tMCAO. (C) NO production was measured (n = 4). Data are presented as the mean ± SEM. P-values were determined by ANOVA followed by the Tukey post hoc test, *P < 0.05; **P < 0.01; ***P < 0.001; and ns, not significant.

Fig 4: Regenerated neurons derived from Nestin+cells in denervated double-transgenic mice (Nestin-creERT2: tdTomato).(A–D) Representative immunofluorescence confocal laser images in freshly intermuscular plexus tissues of gastric of GFAP/β-tubulin/ HuC/D/nNOS (green) and tdTomato-labeled Nestin+ cells (red) in Control, Control+TAM, Sham+TAM, and BAC+TAM groups, the nuclei (blue). Arrows indicate double-positive cells. Nestin+ cells are stained with td-Tomato (red) and express GFAP/β-tubulin/ HuC/D/nNOS (green) after TAM induction and show co-localization, providing evidence that Nestin+ cells can be differentiated into neurons and glial. (E and F) Representative immunofluorescence confocal laser images in freshly intermuscular plexus tissues of gastric of GFAP/β-tubulin/ HuC/D/nNOS (gray), tdTomato-labeled Nestin+ cells (red), and GFP-labeled BMSCs (green) in Sham+BMSCs+TAM and BAC+BMSCs+TAM groups, the nuclei (blue). Control, the control C57 mice; Sham, sham operation group; BAC, benzalkonium chloride treated group; BAC+BMSCs, BAC treated mice transplanted with BMSCs; TAM, tamoxifen.

Fig 5: Proportion of Nestin+/Ngfr+cells differentiated into neurons was significantly higher than that of Nestin+cells.(A) Representative immunofluorescence confocal laser images in freshly intermuscular plexus tissues of gastric of GFAP/β-tubulin/HuC/D/nNOS (green) and tdTomato-labeled Nestin+ cells (red) in BAC+BMSCs+TAM (Nestin-creERT2: tdTomato mice), the nuclei (blue). Arrows indicate double-positive cells. (B) Representative immunofluorescence confocal laser images in freshly intermuscular plexus tissues of gastric of GFAP/β-tubulin/HuC/D/nNOS (red) and GFP-labeled Nestin+/Ngfr+ cells (green) in BAC+BMSCs+TAM (Nestin-creert2 X Ngfr-Dreert2: DTRGFP mice), the nuclei (blue). Arrows indicate double-positive cells. (C and D) Proportion of Nestin+ (red)/Nestin+Ngfr+ (blue) cells differentiated into neurons and glial cells in Control+TAM group and BAC+BMSCs+TAM group. Control, the control C57 mice; Sham, sham operation group; BAC, benzalkonium chloride treated group; TAM, tamoxifen. These results are representative of independent experiments repeated at least 3 times. Results were expressed as mean ± standard deviation. ∗P < .05, ∗∗P < 0.01, NS, no significance.