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

Figure 5.

Figure 5

Representative images and graphs of urothelial proliferation in keratinocyte growth factor (KGF)–pretreated bladders 1 day 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 1 day after cyclophosphamide administration. A and D: Ki-67 staining shows that compared with 6 hours after injury, both the PBS (A) and the KGF (D) groups appear to have more Ki-67+ proliferating urothelial cells 1 day after injury, although the KGF group still appears to have more Ki-67+ cells than the PBS group. Blue indicates DAPI. Dotted lines indicates demarcation between urothelial layer and underlying lamina propria. B and E: Unlike 6 hours after injury, Ki-67 and KRT14 merged staining shows that both PBS-pretreated (B) and KGF-pretreated (E) mice have expansion of KRT14+ cells across the basal layers, many of which are Ki-67+ (arrowheads), although some Ki-67+ cells do not colabel with KRT14 (concave arrowheads). C and F: Ki-67 and KRT5 merged staining shows that most of the Ki-67+ proliferating cells in both the PBS (C) and KGF (F) groups are KRT5+, including in KRT14+ (arrowheads) and KRT14− (concave arrowheads) cells; a rare KRT5− cell is seen in F (arrow). G: Graph confirming higher percentages of proliferating urothelial cells (Ki-67+/DAPI+ cells) in both PBS and KGF groups compared with 6 hours after cyclophosphamide administration and significantly higher percentages of proliferating urothelial cells in the KGF group versus the PBS group 1 day after cyclophosphamide injection (similar to 6 hours after injury). H: Graph showing that the percentages of KRT5+/KRT14− (KRT5 only) proliferating (Ki-67+) urothelial cells are statistically higher in the KGF- versus PBS-pretreated mice, whereas the percentages of KRT14+ and KRT14−/KRT5− (no label) proliferating cells are not statistically different. Data are expressed as means ± SEM (H). N = 9 (3 planes per bladder and 3 bladders per group; H). **P < 0.01. Scale bars = 50 μm (A–F). L, lumen.