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Figure S1.Validation of Utrn-A and Utrn-B antibody specificity using western blots and microdisceted brain capillaries. A. Western blot analysis of lysates containing 50 μg of total protein from mdx brain capillaries were probed with preimmune serum (lane 1), Utrn-A affinity purified antibody (lane 2), and affinity purified antibody preadsorbed with excess Utrn-A peptide (lane 3). Only blot incubated with Utrn-A affinity purified antibody resulted in sharp band for Utrn-A protein. B. Similarly, western blot probed with Utrn-B affinity purified antibody showed sharp Utrn-B protein band (lane 5). Bolts incubated with preimmune serum (lane 4) and affinity purified antibody preadsorbed with excess Utrn-B peptide (lane 6) did not show Utrn-B protein band. C. Validation of microdissection of brain capillaries. Merged image showing isolated brain capillaries immunolabeled with FITC-conjugated lectin antibody (green channel) and co-stained with DAPI (blue channel) as a positive control for brain capillary microdissection. For details of the procedure, see Materials and Methods. Scale bar, 25 μm.

Figure S2.Spatial and temporal changes in Utrn-A and Utrn-B immunolabeling in cultured astrocytes. Astrocytes isolated from mdx mouse were cultured on coverslips coated with poly-L-lysine for 24 and 72 h. They were immunolabeled with Utrn-A and Urn-B antibodies for the spatial and temporal changes in Utrn-A and Utrn-B expression (green channel). A. Astrocytes cultured for 24 h showed distinct Utrn-A immunolabeling in astrocytes end-feet (arrows) and weak staining in the cytoplasm. After 72 h of culture, Utrn-A immunolabeling increased robustly in the astrocytes end-feet, as well as in the cytoplasm. B. Astrocytes cultured for 24 h showed weak Utrn-B immunolabeling in astrocytes end-feet (arrows) and in the cytoplasm; however, after 72 h of culture, there was moderate increase in Utrn-B labeling. Astrocytes were identified by S100-positive labeling (red channel). C. Paxillin, a focal adhesion marker (red channel), was used to double label Utrn-A and Utrn-B in astrocytes focal adhesion points. A well-established co-localization was observed in Utrn-A with paxillin (arrows) in the astrocytes and moderately with Utrn-B (arrows). Scale bar, 100 μm.

Supporting data. Copy number calculation for Utrn-A and -B.

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