Zen blue software version 2

Mouse brains (n = four for control wild type, R26^{GT-Map2k1-GFP/+} or Tg-Cdh5CreER^+/− and n = 4 for mutant R26^{GT-Map2k1-GFP/+}; Tg-Cdh5CreER^+/−) were fixed in formalin, embedded in paraffin, sectioned (seven µm), and hematoxylin/eosin stained. Tissue undergoing immunostaining were flash frozen in OCT and stored at − 80 °C. Seven µm cryo-sections were cut. Primary antibodies used: rabbit anti-mouse COL15A1 (Gift from Dr. Karppinen, University of Oulu, Finland) (1/500) and rat anti-mouse CD31 (1/500) (BD Pharmigen: 553370). Secondary antibodies used: donkey anti-mouse IgG (H + L) Alexa Fluor 488 (Abcam 150105, 1/200); donkey anti-rabbit IgG (H + L) rhodamine red-X (1/500) (Jackson ImmunoResearch 712-295-150). Sections were mounted with DAPI Fluoromount-G (SouthernBiotech). Images of sections were obtained using an Eclipse 80i microscope (Nikon) with a Spot model 25.4 camera and Spot 5.2 software (Diagnostic Instruments). Confocal images were captured using a Zeiss LSM 800 confocal microscope and Zen Blue software version 2.5.