Extended Data Fig. 2: Nissl staining and immunohistochemical analysis of the hippocampal CA1 region and the prefrontal cortex (PFC).

a, Representative images of Nissl staining of the CA1 (up) and PFC (below). b, c, Quantification of the number of cells per standardized area (b) and percentage of ellipsoid cells per area (c) in the CA1 between the experimental groups. d, e, Quantification of the number of cells per standardized area (d) and percentage of ellipsoid cells per area (e) in the PFC between the experimental groups. n = 3. f, h, Representative confocal images of immunofluorescent staining for neurons (NeuN), astrocytes (GFAP), and microglia (IBA1) counterstained with DAPI nuclear stain in CA1 (f) and PFC (h). g, Quantification of GFAP immunoreactivity in hippocampal CA1 region depicting comparable immunoreactivity between OrganEx and 0h WIT group, with a significant increase compared to the other groups. i, j, k, l, Quantification of NeuN immunolabeling intensity (i), number of GFAP+ fragments (j), and number of GFAP+ cells (k) depict similar trends between the groups as seen in the CA1. Microglia number (l) shows comparable results between OrganEx and 0h WIT with different dynamics seen in the ECMO group. n = 3. Scale bars, 50 μm. Data presented are mean ± s.e.m. One-way ANOVA with post-hoc Dunnett’s adjustments was performed. For more detailed information on statistics and reproducibility, see methods. *P < 0.05, **P < 0.01, ***P < 0.001.