EPO induction by HIF-PHIs is attenuated in the ISAM and AnRED mutant mouse lines. (A) Plasma EPO concentrations in the control, ISAM, or AnRED mice at 6 hours after injection of vehicle, GSK360A (50 mg/kg), or roxadustat (200 mg/kg). (B) mRNA expression levels of the Epo, Serpine1, and Egln3 genes in the kidneys or livers of control, ISAM, or AnRED mice at 6 hours after injection of vehicle, GSK360A (50 mg/kg), or roxadustat (200 mg/kg). (C) A semiquantitative RT-PCR analysis was conducted to examine the expression levels of the Epo transgene (Tg^3.3K-EpoE) in the livers of control and ISAM mice at 6 hours after injection of vehicle or GSK360A (50 mg/kg). Based on the band intensities after agarose electrophoresis (representative images of 29- and 33-cycle PCR products are shown in the left panel), the expression levels were estimated (right) after normalization to Actb expression levels. m, size markers. The average mRNA expression levels in the livers of vehicle-treated control mice were normalized to 1 for each gene in panels B-C. Arrows indicate undetectable levels. ∗P < .05 or ∗∗P < .01 calculated by Student t test (control, AnRED, and ISAM in panel A-C) or Tukey-Kramer test (ISAM in panel B).