Fig. 3

mRNA (A) and Western blot (B, C) analyses of the target genes of iron metabolism related miRNAs. Real time PCR was carried out using a SYBR Green protocol. β-actin was used as the housekeeping gene for the normalization of the reaction. The relative gene expression of the control hypothalamus samples was regarded as 1. Real time polymerase chain reactions were carried out in triplicate in each independent experiment. For the WB, the same amount of protein from each lysate was used for the analysis. A mRNA expression levels of iron metabolism related genes FP, TfR1, DMT-1, LF, FTL, and BMPR. B Western blot analysis of the target proteins of iron metabolism FP, TfR1, DMT-1, and FTL. C Optical density analysis of FP, TfR1, DMT-1 and FTL. The columns represent mean values, error bars show the standard deviation (SD) of three independent determinations (n = 3). Asterisk indicates p < 0.05 compared to the control. FP: ferroportin; TfR1: transferrin receptor 1; DMT-1: divalent metal transporter-1; LF: lactoferrin; FTL: ferritin light chain; BMPR: bone morphogenic protein receptor; STZ: streptozotocin