Fig. 2

mRNA (A) and Western blot (B, C) analyses of the target genes of inflammationrelated 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 IL-6, IL-1β, TGFβ, TNF-α, FKN, and fractalkine receptor (CX3CR1). B Western blot analysis of the target proteins of inflammation. C Optical density analysis of the target proteins. 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. IL-6: interleukin-6; IL-1β: interleukin-1β; TGFβ: transforming growth factor β; TNF-α: tumor necrosis factor-α; FKN: fractalkine; CX3CR1: fractalkine receptor; STZ: streptozotocin