Thank you for your advice, nevertheless, I'm still confused that how did researchers get direct evidence to prove these iron-related genes are regulated or associated with iron level within the cell. Isn't it necessary to add iron ion to the liver cell line to confirm the hypothesis ?
Indeed, utilizing RT-qPCR or Immunoblotting can help we understand the gene expression and protein level in hepatocytes. However, isn't it necessary to do these experiments under iron, iron depleted or knockdown conditions?
It would be amazing if there is a condition or protocol to treat iron or iron chelator to the hepatocytes.