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. Author manuscript; available in PMC: 2015 Nov 1.
Published in final edited form as: Hepatology. 2014 Sep 10;60(5):1753–1766. doi: 10.1002/hep.27285

Fig. 7.

Fig. 7

Wnt signaling antagonizes Notch signaling to promote hepatocyte regeneration in the presence of sustained fibrogenic stimulus. (A–F) Compared with the DMSO treatment (A, D), Wnt agonist CHIR-99021 increased the CFP+ cells while downregulating Tg(Tp1:VenusPEST) expression both in the MTZ- (B) and EtOH/MTZ- (E) treated regenerating livers. (C, F) Comparison of the percentage of Tg(Tp1:VenusPEST)high/Alcam+ (MTZ/DMSO, 12.0±4.9%; MTZ/CHIR99021, 10.7±3.2%; EtOH/MTZ/DMSO, 12.0±2.2%; EtOH/MTZ/CHIR99021, 10.8±3.2%), Tg(Tp1:VenusPEST)low/Alcam+(MTZ/DMSO, 25.7±5.2%; MTZ/CHIR99021, 11.8±4.7%; EtOH/MTZ/DMSO, 40±8.3%; EtOH/MTZ/CHIR99021, 30.2±2.3%), Tg(Tp1:VenusPEST)−/Alcam− (MTZ/DMSO, 62.3±9.9%; MTZ/CHIR99021, 77.5±13.6%; EtOH/MTZ/DMSO, 48.0±6.8%; EtOH/MTZ/CHIR99021, 59.0±13.8%), and Tg(Tp1:VenusPEST)−/Alcam−/CFP+ cells (MTZ/DMSO, 13.2±3.2%; MTZ/CHIR99021, 27.5±5.0%; EtOH/MTZ/DMSO, 12.0±1.2%; EtOH/MTZ/CHIR99021, 19.7±6.2%) between the DMSO- vs. Wnt agonist-treated regenerating livers. Wnt agonist treatment significantly increased the total percentage of CFP-positive hepatocytes and Tg(Tp1:VenusPEST)−/Alcam- cells, in which all CFP-positive hepatocytes were derived from, with a decrease in Tg(Tp1:VenusPEST)low/Alcam+ cells both in the MTZ- (C) and EtOH/MTZ- (F) treated regenerating livers. Cells in 5 planes of confocal images from 5 individual larvae were counted. (G) qRT-PCR analysis of numb mRNA in control (7 dpf), MTZ-treated (0 dpa) and MTZ-treated regenerating livers (1 dpa). numb was significantly upregulated in the 1 dpa MTZ-treated regenerating livers. (H, I) Wnt agonist CHIR-99021 upregulated numb expression both in the MTZ- (H) and EtOH/MTZ- (I) treated regenerating livers. n=150 dissected larval livers per condition in three experiments. Asterisks indicate statistical significance: *p<0.05 and **p<0.01. All images are confocal single-plane images (A, B, n=30 larvae per condition in three experiments; D, E, n=15 larvae per condition in three experiments). Scale bars, 20µm. EtOH, ethanol; SD, standard deviation.