HJV-mediated BMP signaling and hepcidin expression are not altered by inhibition of endogenous neogenin expression in Hep3B cells. (A,B) Hep3B cells were transfected with BRE-Luc (A) or Hep-Luc (B) and pRL-TK Renilla luciferase vector, either alone or with increasing amounts of Flag-HJV cDNA, in combination with control siRNA or neogenin siRNA. After 46 hours, cell lysates were analyzed for luciferase activity as in Figure 1B-F. (C) Hep3B cells were transfected with empty vector (pcDNA3) or Flag-HJV cDNA in combination with control siRNA or si-neogenin. After 46 hours, hepcidin relative to RPL19 mRNA levels were quantified by real-time RT-PCR as in Figure 1G,H. Values shown are the means of triplicate measurements plus or minus SD.