Shows the expressional stability of selected reference genes, using NormFinder in hypothalamus and kidney.

<p>NormFinder algorithm is based on model-based approach to calculate overall reference gene stability. Gene with lower standard deviation (SD) is consider the most stable gene. In both tissues Hmbs is the most stable gene while in hypothalamus HPRT and in kidney B2m is the least stable one.</p

Selection of suitable endogenous reference genes for qPCR in kidney and hypothalamus of rats under testosterone influence - Fig 1

<p>(A) Distribution of cycle threshold of 6 different genes in hypothalamus (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0176368#pone.0176368.s001" target="_blank">S1 Table</a>) and kidney (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0176368#pone.0176368.s002" target="_blank">S2 Table</a>); 4 biological and 3 technical replicates were used. Mean Ct value of each gene in all treated groups were described. Lower level of Ct value implies higher expression level as for GAPDH, ACTB, Ppia in both tissue and B2m only in hypothalamus. (B, C) Box plot graphs of Ct values are shown as a square across the box is depicted as the median. The box indicates the 25th and 75th percentiles and the whiskers caps represent the maximum and minimum values.</p

Presents the final optimal list of genes in hypothalamus and kidney.

<p>Presents the final optimal list of genes in hypothalamus and kidney.</p

Presents number of optimal reference gene in hypothalamus (A) and kidney (B) which defined by NormFinder, whereby the lower value of accumulated standard deviation (Acc.SD), identified by red color, indicates the optimal number of reference genes.

<p>Presents number of optimal reference gene in hypothalamus (A) and kidney (B) which defined by NormFinder, whereby the lower value of accumulated standard deviation (Acc.SD), identified by red color, indicates the optimal number of reference genes.</p

Suitable reference genes for both hypothalamus and kidney.

<p>Shows the suitable reference genes for both hypothalamus and kidney were Ppia& Hmbs.</p

Description and efficiency of six reference genes, used for qPCR.

<p>Description and efficiency of six reference genes, used for qPCR.</p

A RNA-Seq Analysis of the Rat Supraoptic Nucleus Transcriptome: Effects of Salt Loading on Gene Expression

<div><p>Magnocellular neurons (MCNs) in the hypothalamo-neurohypophysial system (HNS) are highly specialized to release large amounts of arginine vasopressin (Avp) or oxytocin (Oxt) into the blood stream and play critical roles in the regulation of body fluid homeostasis. The MCNs are osmosensory neurons and are excited by exposure to hypertonic solutions and inhibited by hypotonic solutions. The MCNs respond to systemic hypertonic and hypotonic stimulation with large changes in the expression of their Avp and Oxt genes, and microarray studies have shown that these osmotic perturbations also cause large changes in global gene expression in the HNS. In this paper, we examine gene expression in the rat supraoptic nucleus (SON) under normosmotic and chronic salt-loading SL) conditions by the first time using “new-generation”, RNA sequencing (RNA-Seq) methods. We reliably detect 9,709 genes as present in the SON by RNA-Seq, and 552 of these genes were changed in expression as a result of chronic SL. These genes reflect diverse functions, and 42 of these are involved in either transcriptional or translational processes. In addition, we compare the SON transcriptomes resolved by RNA-Seq methods with the SON transcriptomes determined by Affymetrix microarray methods in rats under the same osmotic conditions, and find that there are 6,466 genes present in the SON that are represented in both data sets, although 1,040 of the expressed genes were found only in the microarray data, and 2,762 of the expressed genes are selectively found in the RNA-Seq data and not the microarray data. These data provide the research community a comprehensive view of the transcriptome in the SON under normosmotic conditions and the changes in specific gene expression evoked by salt loading.</p></div

Transcriptional and Translational Regulator mRNAs decreased in expression by SL.

<p>^aMean Control and Mean SL data are averages of three samples each and are expressed in quantile (log2 (RPKM+2)) units.</p><p>Transcriptional and Translational Regulator mRNAs decreased in expression by SL.</p

Transcriptional and Translational Regulator mRNAs increased in expression by SL.

<p>^aMean Control and Mean SL data are averages of three samples each and are expressed in quantile (log2 (RPKM+2)) units.</p><p>Transcriptional and Translational Regulator mRNAs increased in expression by SL.</p

Pie Chart showing Locations and Types of 552 selected, expressed genes (See columns N and O in S9 Table).

<p>Pie Chart showing Locations and Types of 552 selected, expressed genes (See columns N and O in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0124523#pone.0124523.s018" target="_blank">S9 Table</a>).</p