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. 2026 Sep 3;29(9):117380. doi: 10.1016/j.isci.2026.117380

Figure 2.

Figure 2

Distribution of iPUT in acute mouse hippocampal slices

(A) iPUT accumulation in acute slices in the presence of polyamine uptake inhibitor, BV. Left: representative confocal images of acute hippocampal slices incubated with iPUT (yellow) in the presence of BV. Nuclei are labeled with DAPI (cyan). Scale bars, 500 μm. Right: quantification of mean iPUT fluorescence intensity in slices exposed to iPUT in combination with BV (0, 0.1, and 1 μM) (n = 3 animals).

(B) iPUT distribution in CA1 pyramidal neurons. Top: representative confocal images of acute slices incubated with iPUT (yellow) and labeled for CA1 pyramidal neurons by WFS1 immunofluorescence (magenta). Nuclei are labeled with DAPI (cyan). Images of whole slices (top) and a fragment of the CA1 stratum pyramidale (bottom) are shown. Arrows indicate CA1 pyramidal neurons exhibiting extranuclear accumulation of iPUT within their somata. Scale bars, 500 μm for low-magnification images, 20 μm for high-magnification images. Bottom left: Pearson’s colocalization heatmaps show iPUT distribution between extranuclear (Ex; labeled with WFS1) and intranuclear (In; labeled with DAPI) compartments within individual neurons. Bottom right: quantification of colocalization of iPUT-WFS1 (indicating extranuclear localization) and iPUT-DAPI (indicating intranuclear localization) with Pearson’s correlation coefficient (n = 4 animals).

(C) iPUT distribution in CA1 astrocytes. Top: representative confocal images of acute slices incubated with iPUT (yellow) and labeled for astrocytes by GFAP immunofluorescence (magenta). Nuclei are labeled with DAPI (cyan). Images of whole slices (top) and a fragment of the CA1 stratum radiatum (bottom) are shown. Arrowhead indicates an astrocytic nucleus with prominent iPUT accumulation. Arrows indicate astrocytes exhibiting iPUT accumulation within their processes. Triangle points to an astrocyte with negligible iPUT accumulation. Scale bars, 500 μm for low-magnification images, 20 μm for high-magnification images. Bottom left: Pearson’s colocalization heatmaps show iPUT distribution between extranuclear (Ex; labeled with GFAP) and intranuclear (In; labeled with DAPI) compartments within individual astrocytes. Bottom right: quantification of colocalization of iPUT-GFAP (indicating extranuclear localization) and iPUT-DAPI (indicating intranuclear localization) with Pearson’s correlation coefficient (n = 4 animals). Data information: in (A), (B), and (C), data are presented as mean ± SEM. (A) Statistical analysis: one-way ANOVA, statistically significant effect of BV treatment, F(2,6) = 55.28, p = 0.0001, ∗∗∗p < 0.001, post-hoc Bonferroni test. (B and C) ∗p < 0.05, Student’s t test.