Skip to main content
. 2020 Jan;190(1):108–124. doi: 10.1016/j.ajpath.2019.09.015

Figure 7.

Figure 7

Representative images and graphs of urothelial proliferation in keratinocyte growth factor (KGF)–pretreated bladders 3 days after cyclophosphamide injection. A–F: Triple-label immunofluorescence for Ki-67 (red), KRT14 (white), and KRT5 (green) in PBS-pretreated (A–C) and KGF-pretreated (D–F) bladder sections 3 days after cyclophosphamide administration. A and D: Ki-67 staining shows that compared with 6 hours and 1 day after injury, both the PBS group (A) and the KGF group (D) appear to have more Ki-67+ proliferating nuclei 3 days after injury, although now the PBS group has many more Ki-67+ cells than the KGF group, including three to four rows of proliferating cells in the former versus one to two in the latter. Blue indicates DAPI. Dotted lines indicate demarcation between urothelial layer and underlying lamina propria. B and E: Ki-67 and KRT5 merged staining shows that both PBS-pretreated mice (B) and KGF-pretreated mice (E) have expansion of KRT14+ cells covering the basal layers and into more superficial layers (two to four total layers in the former and one to three layers in the latter), many of which are Ki-67+ (arrowheads), although some Ki-67+ cells do not colabel with KRT14 (arrows). C and F: Ki-67 and KRT5 merged staining shows that most of the Ki-67+ proliferating urothelial cells from both the PBS group (C) and the KGF group (F) are KRT5+, including in KRT14+ (arrowheads) and the few KRT14− (arrows) cells. G: Graph showing higher percentages of proliferating cells (Ki-67+/DAPI+ cells) in both the PBS and KGF groups compared with 6 hours and 1 day after injury and significantly higher percentages of proliferating urothelial cells in the PBS group versus the KGF group 3 days after cyclophosphamide injection (unlike at 6 hours and 1 day after injury). H: Graph showing that the percentages of KRT14+ proliferating (Ki-67+) urothelial cells are statistically higher in the PBS- versus KGF-pretreated mice, whereas the percentages of KRT5+/KRT14− (KRT5 only) and KRT14−/KRT5− (no label) proliferating cells are not statistically different. Data are expressed as means ± SEM (H). N = 6 (2 planes per bladder and 3 bladders per group; H). **P < 0.01, ***P < 0.001. Scale bars = 50 μm (A–F). L, lumen.