Confocal microscopy and double immunostaining of Egr-1 and PECAM in the heart of VEGF- and EGF-treated mice.

Mice received intraperitoneal injections of normal saline, VEGF, or EGF. Hearts were removed 1 hour later. Tissue sections were double immunostained with a rabbit polyclonal anti-Egr-1 antibody and a rat monoclonal anti-CD31 antibody, washed in PBS, and incubated with Cy3-conjugated goat antirabbit IgG and FITC-conjugated goat antirat IgG. (Top) Low-power view of optical sections through the myocardium of a normal saline-treated animal (A-C), a VEGF-treated animal (D-F), and an EGF-treated animal (G-I). Egr-1 immunofluorescence is shown on the left (A, D, G), and PECAM immunofluorescence is shown in the middle (B, E, H). The 2 images are combined on the right (C, F, I). Bar, 100 μm. (Bottom) High-power view of optical sections through the myocardium of VEGF-treated animal (A-C) and EGF-treated animal (D-F). Egr-1 immunofluorescence is shown on the left (A, D), and PECAM immunofluorescence is shown in the middle (B, E). The 2 images are combined on the right (C, F). Bar, 25 μm.