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. 2008 Oct 10;295(6):H2242–H2249. doi: 10.1152/ajpheart.00587.2008

Fig. 2.

Fig. 2.

Dual fluorescence combining TNF-α with markers for endothelial cells [von willebrand factor (vWF)] and vascular smooth muscle (α-actin) with the use of specific antibodies, followed by fluorescent-labeled secondary antibodies. A, B, and C: dual labeling of TNF-α (red) and α-actin (green) in sham control (CTL) mouse heart tissue. D, E, and F: dual labeling of TNF-α (red) and α-actin (green) in I/R mouse heart tissue. The pink arrow shows the staining of TNF-α (red), and the blue arrow shows the colocalization of TNF-α and vascular smooth muscle cells (yellow). G, H, and J: dual labeling of TNF-α (red) and vWF (green) in I/R mouse heart tissue. The white arrows in I and K show the specific vWF staining with absence of TNF-α staining. L and M: negative CTL. The purple arrows show an absence of staining in vessels with the secondary antibodies. N shows nuclear staining with 4,6-diamidino-2-phenylindole (blue) in I/R mouse heart tissue. Magnification ×40. Data shown are representative of 4 separate experiments.