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Treatment with sex hormones affects hepatocyte proliferation during NAHCC progression in adult gMX2 and hMX3 male fish. WT ± Dox, gMX2 ± Dox, and hMX3 ± Dox (6 mpf) groups were treated with E2 and 11-KT for 90 days. (A) Representative gross observations (Scale bar: 2.5 mm) and (B) liver histology of WT ± Dox, gMX2 ± Dox, and hMX3 ± Dox after no treatment (control) and treatment with E2 and 11-KT. Scale bar: 100 μm. (C) Percentage of NAHCC phenotypes observed from the liver histology in (B). Scale bar: 100 μm. (D) PCNA staining of liver slices of male zebrafish in distinct experimental groups. Scale bars, 100 μm. Insets, Scale bars, 25 μm. (E) Percentage of hepatocyte proliferation in the male zebrafish in discrete treatment groups (n = 5−10). (F) Western blot analysis in the different treatments of male hMX3 ± Dox livers. Statistically significant differences from the controls are indicated by * (p < 0.05), and ** (p < 0.01).
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