RT-qPCR of selected genes.

<p>Gene expression was determined by RT-qPCR in eosinophils isolated from subjects with asthma and healthy controls. Results were normalized to <i>β-actin</i> and expressed as fold change compared with samples from healthy controls. Results are presented as means and SEM. *p<0.05; **p<0.01 (2-tailed non parametric Mann-Whitney).</p

Eosinophils from asthmatic subjects display a similar transcriptional profile as eosinophils from other hypereosinophilic conditions.

<p>Scatterplots are based on fold changes of selected genes that are differentially expressed at least in one hypereosinophilic disease (p< 0.01). Diagonal: estimation of the density of log fold changes (lfc) for each variable. Upper elements: estimation of the bivariate density of each couple of variables. In red: genes with a lfc changed to the same direction in all the conditions. In black: genes with a lfc changed to the opposite direction in at least one couple of variables. With a cut-off for the lfc of 0.15, more than 95% of the probe sets have identical changes. Ast, asthma; Asp, pulmonary aspergillosis; Der, dermatological disease; Par, parasitosis.</p

Circulating eosinophils in asthma differ in their gene expression profile when compared to healthy subjects.

<p>Heat map of hierarchical clustering of the top expressed genes of circulating eosinophils from subjects with asthma (<i>n</i> = 4) <i>vs</i> healthy controls (<i>n</i> = 3). The horizontal dendrogram represents the relationship between asthmatic and healthy subjects. The vertical dendrogram represents the relationship between the expression levels of each gene across all the samples. Over-expressed genes are shown in red and under-expressed genes are depicted in green.</p

Gene ontology categories with an enrichment score p<0.01 and > 10% of gene concordance identified using the DAVID database.

<p>Gene ontology categories with an enrichment score p<0.01 and > 10% of gene concordance identified using the DAVID database.</p

Eosinophil isolation by FACS generates high quality RNA.

<p><b>(A)</b> Flow cytometry gating strategy for the identification of eosinophils. Eosinophils were identified among a granulocyte suspension (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0141740#sec002" target="_blank">Methods</a>) as a CD16 fluorescence negative cell population <b>(B)</b> Purity of sorted eosinophils as assessed by cytospin preparation was close to 100%. <b>(C)</b> Bioanalyzer RNA profile with RNA integrity number (RIN) of an eosinophil sample. All RNA samples included in the expression analysis had a RIN > 8.</p

Subject characteristics.

<p>Subject characteristics.</p