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

نویسندگان

  • Kory R. Johnson
  • C. C. T. Hindmarch
  • Yasmmyn D. Salinas
  • YiJun Shi
  • Michael Greenwood
  • See Ziau Hoe
  • David Murphy
  • Harold Gainer
چکیده

There are a number of errors in the captions for S1 to S9 Fig. in the Supporting Information. Please see the complete, correct S1 to S9 Fig. captions here. S1 Fig. Micrograph illustrating magnocellular neurons (MCNs) in a salt loaded SON mounted on a PENmembrane frame slide and visualized by a Arcturus XT laser capture microscope. Large blood vessel adjacent to the SON is shown in the upper left corner. Scale line is 100um. S2 Fig. RNA-Seq Analysis Workflow. (A) Overview of analysis steps performed. Step 1, "Sequence Adaptor Clipping”. Read pairs, 101bp in length, were adaptor clipped via FASTQ/A Clipper (http://hannonlab.cshl.edu). Broken read pairs as a result of clipping were discarded. Step 2, “Sequence Quality Inspection”. Intact read pairs remaining post-clipping were import into CLCbio and the “Create Sequencing QC Report” tool used to generate one report per sample. These reports, each containing per-sequence and per-base quality statistics, were individually inspected and cross-compared to define universal hard-trimming rules to be applied to all samples. Step 3, “Sequence Quality Trimming & Filtering”. The CLCbio “Trim Sequences” tool was used to hard-remove the first 15nt from the 5’ end and the last 1nt from the 3’ end of each read pair. The tool was also used to dynamically-trim away nucleotides having a call accuracy rate less than 95%. Read pairs having at least one sequence containing more than two ambiguities were also discarded as part of this Step as were read pairs having at least one sequence with a post trimmed length less than 15 nucleotides. Step 4, “Sequence Alignment and Enumeration”. The CLCbio “RNA-Seq Analysis” tool was used to align read pairs to the Rat Genome (RN5) by sample using default parameters. Output provided by the tool included a Reads per kilo base per million (RPKM) expression value for 26,313 genes. Step 5, “RPKM Expression Pedelstalling”. Output from Step 4 was imported into R (http://www.r-project.org/) and a value of two added to each RPKM expression value per sample. Step 6, “RPKM Expression Transformation”. Pedestalled values from Step 5 (RPKM+2) were Log2 transformed using standard commands in R then filtered to keep only those genes having a post-transformed expression value (Log2(RPKM+2))>1 for at least one sample. Step 7, “RPKM Expression Normalization”. Transformed values from Step 5 were quantile normalized using standard commands in R. Step 8, “Exploratory Analysis”. Normalized values from Step 7 (Quantile(Log2 (RPKM+2))) were interrogated in R by Tukey box plot, covariance-based principal component analysis (PCA) scatter plot and Pearson correlation-based heat map to confirm absence of outliers. Step 9, “Noise Analysis”. For each gene, the coefficient of variation (CV) and mean expression was calculated by sample class using standard commands in R then modeled by sample class using the lowess() command. Step 10, “Confidence Criterion Selection”. Lowess fits from Step 9 were visually inspected to define the mean expression value across sample

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

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

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 ...

متن کامل

I-13: Transcriptome Dynamics of Human and Mouse Preimplantation Embryos Revealed by Single Cell RNA-Sequencing

Background: Mammalian preimplantation development is a complex process involving dramatic changes in the transcriptional architecture. However, it is still unclear about the crucial transcriptional network and key hub genes that regulate the proceeding of preimplantation embryos. Materials and Methods: Through single-cell RNAsequencing (RNA-seq) of both human and mouse preimplantation embryos, ...

متن کامل

Unsupervised Network Analysis of the Plastic Supraoptic Nucleus Transcriptome Predicts Caprin2 Regulatory Interactions

The supraoptic nucleus (SON) is a group of neurons in the hypothalamus responsible for the synthesis and secretion of the peptide hormones vasopressin and oxytocin. Following physiological cues, such as dehydration, salt-loading and lactation, the SON undergoes a function related plasticity that we have previously described in the rat at the transcriptome level. Using the unsupervised graphical...

متن کامل

Clustering of Short Read Sequences for de novo Transcriptome Assembly

Given the importance of transcriptome analysis in various biological studies and considering thevast amount of whole transcriptome sequencing data, it seems necessary to develop analgorithm to assemble transcriptome data. In this study we propose an algorithm fortranscriptome assembly in the absence of a reference genome. First, the contiguous sequencesare generated using de Bruijn graph with d...

متن کامل

Network-based transcriptome analysis in salt tolerant and salt sensitive maize (Zea mays L.) genotypes

Identification of genes involved in salinity stress tolerance provides deeper insight into molecular mechanisms underlying salinity tolerance in maize. The present study was conducted in the faculty of agriculture of Urmia university, Iran, in 2018, with the aim of identifying genetic differences between two maize genotypes in tolerance to salinity stress, and the results of gene expression wer...

متن کامل

Activation of Wnt signaling reduces high-glucose mediated damages on skin fibroblast cells

Objective(s): High-glucose (HG) stress, a mimic of diabetes mellitus (DM) in culture cells, alters expression of a large number of genes including Wnt and NF-κB signaling-related genes; however, the role of Wnt signaling during HG-mediated fibroblast damage and the relationship between Wnt and NF-κB signaling have not been understood. In this study, we aimed to investigate the ffects of Wnt sig...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:

دوره 10  شماره 

صفحات  -

تاریخ انتشار 2015