Immunohistochemical analyses were performed as previously described.7 (link),21) (link) Briefly, at the indicated period after aneurysm induction, 5-μm-thick frozen sections were prepared. After blocking with 3% donkey serum (Jackson ImmunoResearch, Baltimore, MD, USA), slices were incubated with primary antibodies followed by incubation with secondary antibodies conjugated with a fluorescence dye (Jackson ImmunoResearch). Finally, fluorescent images were acquired on a confocal fluorescence microscope system (Lsm-710, Carl Zeiss Microscopy GmBH, Gottingen, Germany).
The following primary antibodies were used: anti-actin, α-smooth muscle-Cy3 antibody produced in mouse (#C6198, SIGMA, St. Louis, MO, USA), purified rat anti-mouse CD31 (#550274, BD Pharmingen, Franklin Lakes, NJ, USA).
The following primary antibodies were used: anti-actin, α-smooth muscle-Cy3 antibody produced in mouse (#C6198, SIGMA, St. Louis, MO, USA), purified rat anti-mouse CD31 (#550274, BD Pharmingen, Franklin Lakes, NJ, USA).
