RT-qPCR Ct values for reference genes.

<p>Ct values for each reference gene in all Hyperaccumulating ecotype of <i>S. alfredii</i> samples. A line across the box depicts the median. The box indicates the 25% and 75% percentiles, whiskers caps represent the maximum and minimum values, dots represent outliers.</p

Pairwise variation (V) calculated by geNorm to determine the optimal number of reference genes.

<p>The average pairwise variations (Vn/Vn+1) were analyzed to measure the effect of adding additional reference gene on the RT-qPCR normalization for all samples.</p

Expression stability (<i>M</i>) of ten reference genes across all 31 samples calculated by geNorm.

<p>(a) all 31 samples, (b) different tissue/organs, (c) Cd²⁺ treatment, (d) Pb²⁺ treatment, (e) Zn²⁺ treatment, (f) Cu²⁺ treatment.</p

Confirmation of primer specificity and amplicon size.

<p>(a) Melting curves of ten candidate reference genes and one target gene showing single peaks. (b) Agarose gel (2%) showing specific RT-qPCR products of the expected size for each genes. M1 and M2 represent 2000bp and 500bp DNA marker respectively.</p

Example for adding a term to ESND.

<p>Example for adding a term to ESND.</p

Screenshot of the term “Exon” in ESND (http://esnd.big.ac.cn/index.php/Exon).

<p>Screenshot of the term “Exon” in ESND (http://esnd.big.ac.cn/index.php/Exon).</p