Figure 3.

Astrocytic cell density did not change in the dentate gyrus of Usp9x^−/Y; Emx1-Cre mice. Co-immunofluorescence labeling and confocal microscopy was performed on hippocampal sections of Usp9x^loxP/Y (A–D) and Usp9x^−/Y; Emx1-Cre (E–H) at P14. Cell nuclei were labeled with DAPI (A, E). Astrocytes were defined as cells expressing both s100β (red in B and F) and GFAP (green in C and G). The merged panels are shown in (D, H). The insets reveal a higher magnification view of the boxed region showing astrocytes (arrowheads). Quantification of labeled cells was performed within the hilar region of the dentate gyrus. There were significantly fewer total numbers of GFAP⁺/s100β⁺ astrocytes in the mutant mice compared to controls (I). Normalized cell counts relative to the volume of the hilar region revealed no significant changes in astrocytes per mm³ within the mutant compared to controls (J). (ns) Not significant, **p < 0.001, t-test. Scale bar in (A): (A-H) – 150 µm; (A’): (A’-H’) −10 µm.