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. Author manuscript; available in PMC: 2019 Dec 28.
Published in final edited form as: Cell Rep. 2019 Sep 24;28(13):3338–3352.e6. doi: 10.1016/j.celrep.2019.08.063

Figure 6. Inhibition of Wnt/β-Catenin Signaling Transiently Reduces Stemness and Promotes Differentiation in Human BCs.

Figure 6.

(A–F) BCIs in ALI culture for up to 4 weeks. Human recombinant DKK1 (DKK1) was used to inhibit Wnt/β-catenin signaling starting at ALI day 7 (D7).

(A) Confocal imaging of specimens stained for Ac.-α-tubulin (MCCs, blue), CC10 (Club cells, green), and Actin (cell membranes, magenta) shows moderately increased MCC and Club cell numbers after DKK1 treatment.

(B) DKK1 leads to a transient decrease in BCs but does not lead to loss of ΔN-TP63+ (green) cells. Nuclei (DAPI, blue).

(C) Quantification from (A) and (B), respectively. Mann Whitney test, not significant, ns (p > 0.05); ***p ≤ 0.001.

(D) Quantitative real-time PCR expression levels are depicted relative to stage controls. DKK1 increases expression of MCC (FOXJ1, MCIDAS) and to a lesser extent Club cell (SCGB1A1) markers but without reduction of BC markers (ΔN-TP63, KRT5). Student’s t test, not significant, ns (p > 0.05); *p ≤ 0.05; **p ≤ 0.01.

(E and F) Optical orthogonal sections of confocal images after staining for BC markers ΔN-TP63 (green, in E) and KRT5 (green, in F).

Related to Figure S5.