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Fig. 4. Immunohistochemical and immunoblot analyses showing increased expression of HIF1 ${alpha}$ in the affected Sox17^+/--Sox18^-/- livers at P7. (A,B) Anti-HIF1 ${alpha}$ staining of Sox18^-/- (+/+) and affected Sox17^+/--Sox18^-/- (+/-) livers at P7. White and black arrowheads indicates HIF1 ${alpha}$ -positive and HIF1 ${alpha}$ -negative hepatocytes, respectively. (B) Magnified images of respective panels in A. (C) Immunoblot analysis of Sox18^-/- (left), mild Sox17^+/--Sox18^-/- (middle) and severe Sox17^+/--Sox18^-/- (right) livers at P7 (two independent samples in each group), showing increased expression of HIF1 ${alpha}$ in the Sox17^+/--Sox18^-/- livers in a phenotype-dependent manner. Numbers indicate relative HIF1 ${alpha}$ expression levels normalized to ACTIN level ± standard error (amount of HIF1 ${alpha}$ in the Sox18^-/- liver is set as 1.0; n=3). Bar, 50 µm.

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