Figure 4. Validation of novel brain pericyte markers by in situ hybridization and immunohistochemistry.
RNA in situ hybridization of 2-month old C57BL6 mouse cerebral cortex. (A) Vitronectin (Vtn, in red) is expressed in PC, identified by Pdgfrb expression (in green). While Pdgfrb expression is restricted to the cell body, Vtn can be found also in PC processes. Arrowhead points to a PC, arrow points to a vascular smooth muscle cell. In magnified inset is seen an overlap of Pdgfrb and Vtn expression in a PC. Cell nuclei are visualized by DAPI (in blue) and EC are visualized by Pecam1 expression (in white). (B) Interferon induced transmembrane protein 1 (Ifitm1, in red) is expressed in PC, identified by Pdgfrb expression (in green). In inserts, cell nuclei are encircled with dotted lines. Arrow points to a PC nucleus, arrowhead points to an EC nucleus. Cell nuclei are visualized by DAPI (in blue) and EC are visualized by Pecam1 expression (in white). (C) Immunohistochemical detection of vitronectin (in red) in PC (in green, CD13). The higher magnification image (C’) shows a strong expression of vitronectin in PC cell body and processes. The scale bars in top and lower panels are 20 μm (A,B), 50 μm (C) and 10 μm (A,B and C’), respectively.