HJV overexpression affects iron homeostasis in mice. (A) Serum iron levels (control PBS, n = 8; control AAV-HJV, n = 5; Alk2^fl/fl;Alb-Cre PBS, n = 6; Alk2^fl/fl;Alb-Cre AAV-HJV, n = 5; Alk3^fl/fl;Alb-Cre PBS, n = 5; and Alk3^fl/fl;Alb-Cre AAV-HJV, n = 6) and (B) transferrin saturation were determined to analyze systemic iron homeostasis (control PBS, n = 9; control AAV-HJV, n = 5; Alk2^fl/fl;Alb-Cre PBS, n = 6; Alk2^fl/fl;Alb-Cre AAV-HJV, n = 5; Alk3^fl/fl;Alb-Cre PBS, n = 5; and Alk3^fl/fl;Alb-Cre AAV-HJV, n = 6). (C) Liver iron content was measured to determine tissue iron retention (control PBS, n = 11; control AAV-HJV, n = 9; Alk2^fl/fl;Alb-Cre PBS, n = 7; Alk2^fl/fl;Alb-Cre AAV-HJV, n = 5; Alk3^fl/fl;Alb-Cre PBS, n = 6; and Alk3^fl/fl;Alb-Cre AAV-HJV, n = 6). (D) Hepatic Bmp6 mRNA levels were determined by qRT-PCR. 18S ribosomal RNA was used as an internal control and the average of control mice treated with PBS was set to 1 (control PBS, n = 10; control AAV-HJV, n = 10; Alk2^fl/fl;Alb-Cre PBS, n = 7; Alk2^fl/fl;Alb-Cre AAV-HJV, n = 5; Alk3^fl/fl;Alb-Cre PBS, n = 6; and Alk3^fl/fl;Alb-Cre AAV-HJV, n = 6). Data are presented as box plots with minimum to maximum whiskers and medians. Significances were presented relative to the indicated control with ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001.