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. 2020 Jun 28;12(7):1712. doi: 10.3390/cancers12071712

Figure 2.

Figure 2

Differentiation markers of urothelium after BBN treatment and vitamin A rich diet. (A–D) Keratin 20 (KRT20) immunofluorescence. KRT20 labelling (green) is strong in the umbrella cells of NT and VitA groups. In BBN and BBN + VitA groups, two different areas are observed. (C,E) In some areas, the apical cytoplasm of individual superficial cells is positive and (D,F) in other areas, superficial, intermediate, and even basal cells are positive. White line depicts the location of basal lamina. L, lumen. Scale bars = 50 μm. Uroplakin immunohistochemistry: (G,H) Uroplakin labelling (brown) is prominent in all superficial urothelial cells of NT and VitA groups; (I,J) only some superficial urothelial cells of BBN and BBN + VitA groups are uroplakin positive. Scale bars = 50 μm. Scanning electron microscopy: Apical plasma membrane of umbrella cells shows (K,L) urothelial plaques (thin arrows) in NT and VitA groups and (M,N) ropy ridges (thin arrows) and microvilli (thick arrows) in BBN and BBN + VitA groups. Scale bars = 5 μm. Four-point scale evaluation of differentiation markers in NT (n = 6), VitA (n = 6), BBN (n = 12), and BBN + VitA (n = 12) groups: (O,P) KRT20 and uroplakin labelling as observed in superficial (SC), intermediate (IC), and basal (BC) urothelial cells; (Q) urothelial apical surface as observed by scanning electron microscope showing percentage of apical surface covered by urothelial plaques, ropy ridges, and microvilli.