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

Supplemental Figure S8.

Supplemental Figure S8

Representative images of Ki-67 and urothelial marker staining in keratinocyte growth factor (KGF)–pretreated bladders from 28 days after cyclophosphamide injection. A and F: Immunofluorescence (IF) for KRT20 (red) shows regions of incomplete staining along the lumenal surface of PBS-pretreated mice (arrowhead), whereas KGF-pretreated mice have strong, continuous signal (F) 28 days after injury. BD: Triple-label IF for Ki-67 (red), uroplakin 3a (UPK3; white), and KRT5 (green) in a section adjacent to A shows that PBS-pretreated mice still have proliferating cells along the basal layer that colabel for KRT5 and not UPK3 (arrowheads), 28 days after injury. E: Co-IF for Ki-67 (red) and KRT14 (white) in a section adjacent to D shows that PBS-pretreated mice continue to have many KRT14+ cells along the basal layer, many of which are proliferating (concave arrowheads) 28 days after injury. GI: Triple-label IF for Ki-67 (red), UPK3 (white), and KRT5 (green) in a section adjacent to F shows that KGF-pretreated mice have no proliferating urothelial cells, including in UPK3+ and KRT5+ cells. J: Co-IF for Ki-67 (red) and KRT14 (white) in a section adjacent to I confirms that KGF-pretreated urothelial cells are not proliferating, although small patches of persistent KRT14+ cells are apparent along the basal layer (arrows) 28 days after injury. A, B, F, and G: Blue indicates DAPI. Scale bar = 50 μm (AJ). L, lumen.