FIG 3.

Disruption of all three PHD isoforms in the liver causes steatosis and growth retardation. (A) Immunoblots for HIF1α and HIF2α in nuclear extracts from the livers of P123-LKO mice at the indicated ages. Nup62 was used as an internal control. (B) Expression levels of Bnip3, Vegfa, Fasn, and Scd1 mRNAs in the livers of PHD mutant mice were analyzed by RT-qPCR. β-Actin was used as an internal control. The expression levels of control mice were set as 1.0. The data are means and SD (n = 3 for each group). (C) Lipid droplets in the livers of the PHD mutant mice at 7 weeks of age were stained with Nile red (red). Hoechst 33342 was used for nuclear staining (blue). (D) Glycogen deposition in the livers of PHD mutant mice at 8 weeks of age was detected by PAS staining (arrows). Bars, 500 μm (C) and 100 μm (D). (E) The glycogen concentration was measured in the livers of the control and P23-LKO mice at 8 weeks of age. The data are means and SD (n = 4 for each group). (F) Changes in body weight between 2 and 5 weeks after birth in P123-LKO and control mice. Error bars indicate SD (n = 3 for each group). (G) Kaplan-Meier survival analysis of the indicated genotype mice (n = 20 for each group). More than half of the P123-LKO mice died by 10 weeks after birth. *, P < 0.01 compared with the control mice (B, E, and F).