Fig. S1. Astrocyte population in the P4 rat nucleus hypoglossus and spinal cord. (A) Example of a confocal image (20×) of hypoglossal nuclei labeled with GFAP (fibrous astrocyte marker; green) and Hoechst 33342 (blue). The inset (higher magnification) shows how GFAP labeling is limited to the central canal profile. (B) Example of GFAP (green) and S100B (protoplasmic astrocytic marker; red) immunoreactivity of the spinal cord ventral horn (fluorescence microscopy image, l0×). (C) Histological example of GFAP (green) and DAPI (blue) staining of cell in the rat spinal cord (fluorescence microscopy image, 5×).

Fig. S2. Immunostaining of motoneurons and astrocytes in the nucleus hypoglossus of P4 rat. (A) Example of a confocal images (l0×) of hypoglossal nuclei labeled with SMI 32 (motoneuron marker; green) and Hoechst 33342 (blue). Inset shows schematic location of nuclei in brainstem section. (B) Higher magnification (20×) of the example in A, showing the region of interest (ROI: white box; 100 000 µm2) used for cell counting. (C) Histological example of S100B (green; astrocytes) and Hoechst 33342 (blue) staining.

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