Figure 5.

In Brainstem Facial Nucleus NCX2 Transcript and Protein Levels Were Downregulated in the Early Phase of SMA Pathology, While They Were Upregulated in the Late Phase of the Disease, Particularly in NeuN-Positive Cells

(A and B) Real-time PCR (A) and western blot analysis (B) for NCX2 mRNA and protein in the brainstem of WT (white columns) and SMA (black) mice, five (P5) and 12 (P12) days after birth showing that NCX2 expression is inversely correlated to miR-206 modulation. (C) Confocal images showing the double labeling of NCX2 (red) and NeuN (green) in the facial nuclei of brainstem slices (40 μm) of WT+NT-miR (a–d); WT+miR-206 (e–h); SMA+NT-miR (i–l), and SMA+miR-206 (m–p) animals at P12. Nuclei were labeled with Hoechst (blue). The merge panels (d, h, l, and p) showed the co-localization of NCX2 in neurons as indicated by arrows. Scale bar, 50 μm. (D) Quantification of NCX2 fluorescence in neurons of WT+NT-miR (white column), WT+miR-206 (light gray), SMA+NT-miR (black), and SMA+miR-206 (dark gray) animals at P12. *p < 0.05; **p < 0.005; ***p < 0.001 by one-way ANOVA analysis followed by Bonferroni test. Each column represents the mean ± SEM (n = 3/5).