Figure 2
Expression of BMAL1 in white matter cells. Sham or SCI mice were killed at ZT 3–5. Co-immunostaining for BMAL1 and the indicated cell type-specific markers were performed on longitudinal (coronal) sections that included the injury epicenter and penumbral regions; nuclei were counterstained with Hoechst-33258. The specificity of the BMAL1 signal was validated by showing its loss in Bmal1−/− spinal cord tissue or after replacing the anti-BMAL1 antibody with a control IgG (Supplementary Fig. S1). As acute cell loss in the injury epicenter may affect identification of BMAL1-expressing cells, all images from SCI animals depict the penumbral region (0.5–1 mm rostral from the injury epicenter). (a) Nuclear BMAL1 signal is present in numerous cells throughout the grey and white matter with similar patterns in sham and SCI mice. The sectioning level is indicated by the red line. The images depict the ventral horn grey matter and the ventrolateral white matter after co-staining for BMAL1 and the OL marker CC1. Position of high power images that are shown in (b) is indicated. (b) High power images of BMAL+ white matter cells that were co-stained for the OL marker CC1, the astrocyte marker GFAP and the EC marker PECAM1. Arrows point double positive cells. All co-immunostainings were done on adjacent sections that were cut as in (a); low power images are shown in Supplementary Figs. S2–S5. The images represent analysis of 3 sham and 3 SCI WT female mice.
