Figure 1
Effect of metformin on BMP6-induced hepcidin gene expression in WT and AMPKLiv KO mice.(A) Q-PCR analysis showing hepcidin mRNA levels in mouse primary hepatocytes treated with BMP6 (10 nM) or/and metformin (from 0.25 to 1 mM) for 24 h. RNA extraction and real-time quantification of transcripts were performed as described14. mRNA expression was calculated using the ΔΔCt method and normalized to the expression of cyclophilin A. Hepcidin gene expression in BMP6 treated hepatocytes, with or without metformin, is expressed as-fold hepcidin expression of that in non-treated hepatocytes. Error bars represent SD for n = 3 samples in each group. Statistical significance is indicated by * (*p < 0.05, **p < 0.005, ***p < 0.0005 as compared to the BMP6 condition). A typical representative experiment is shown. Similar results were confirmed in three separate experiments. (B) Immunoblots showing pAMPK/AMPK and pACC/ACC in hepatocyte extracts prepared as previously described, treated either with 10 nM BMP6, 1 mM metformin or both.
