Hjv^−/− hepatocytes exhibit impaired Smad signaling in response to BMP6. Primary hepatocytes were isolated from livers of wt and Hjv^−/− mice. The cells were cultured in serum-free media and left untreated or pretreated overnight with 30 μM holo-Tf. On the next day, the cells were supplemented (or not) without previous wash with 20 ng/mL murine IL-6, 5 ng/mL human BMP6, or 5 ng/mL human/mouse/rat BMP2, alone or in combinations, as indicated. The incubation was terminated after 4 hours and the cells were harvested and lysed. Hepatocyte RNA and protein extracts were analyzed by qPCR and western blotting, respectively. (A) Expression of Hamp mRNA; (B) levels of pStat3, Stat3, pSmad5, and Smad5; (C) Id1 mRNA; (D) Socs3 mRNA. All data are presented as the mean plus or minus SEM. Statistical analysis was performed by 2-way ANOVA. Statistically significant differences (P < .05) across genotypes are indicated by an asterisk (*).