Figure 5.

Dual fluorescence and peroxidase immunohistochemistry in the regenerating skin marginal to the wound after 5 days of injury. Sections are stained with Ki67–Alexa 594 (A), BrdU–Alexa 488 (B), merged images of Ki67–Alexa 594 and BrdU–Alexa 488 to see the colocalization (C), and BrdU-peroxidase-DAB counterstained with hematoxylin (D). Red arrows indicate S-phase cells (BrdU+/Ki67+ cells) and blue arrows non-S-phase cells (BrdU−/Ki67+). Ki67 and/or BrdU stainings were observed only in the stratum basale and not in the thickened stratum spinosum (*: asterisk). Note the high incidence of S-phase cells (BrdU+/Ki67+ cells) in the stratum basale marginal to the wound. Margin of wound is shown in a broken line. Squared area of D shows the area of the photographed region of fluorescent data of A to C. Bars = 20 µm.