Modulation of RET-independent genes.

<p>Histogram bar graph showing the relative transcript levels of genes that, regardless of treatment with GDNF and GFRα1, are differently modulated in PBMCs from healthy donors and HSCR patients. The amounts of mRNA either in freshly purified (left part of the graph) or treated (right part of the graph) of PBMCs of healthy donors (white bars) and HSCR patients (black bars) were detected by Taqman Low Density Array (TLDA) card.</p

Correlation between the RET receptor expression and RET transcripts on four different cell lines.

<p>(A) Flow cytometric histogram graphs showing the MFI levels of expression of RET receptor (black line) of 4 different cell lines. Gray shaded histograms represent the isotype controls. (B) Histogram bar graph showing the number of RET mRNA copies produced by the same cell lines displayed in panel A and analyzed within the same time-frame in culture. Values are normalized on SK-N-MC cell line of one experiment and are reported as fold increased in expression (2^−ΔCt) as mean of three independent experiments. Of note, the level of RET receptor expressed on cell membrane significantly correlated with the amount of RET transcripts, as assessed by the Spearman rank test for correlation.</p

Expression of RET receptor on immune cells from HSCR patients associated with a single nucleotide polymorphism at exon 2.

<p>Summary graph of statistical dot plots showing MFI values of RET receptor expressed on lymphocytes and monocytes from 50 HSCR patients with medians (horizontal black bars) either in the absence (A) or in the presence (B) of an association analysis stratification based on the genotype at the exon 2 of RET gene (SNP rs1800858). We did not detect any statistically significant association between the phenotypic distribution of RET receptor on immune cells with RET genotype at exon 2 SNP.</p

Expression of RET receptor on immune cells from HSCR patients associated with pathogenic mutations of RET gene.

<p>Summary graph of statistical histogram bars (upper panel) and dot plots (lower panel) showing MFI values of RET expressed on monocytes, T, B and NK lymphocytes from 46 HSCR patients stratified on the basis of individuals either carrying (gray histogram bars in upper panel and black symbols in the lower panel) or not carrying (empty histogram bars in upper panel and empty symbols in the lower panel) pathogenic RET mutation. 4 HSCR patients were excluded from the analysis because they were carrying mutations of the RET gene with uncertain effects.</p

Levels (pg/ml) and standard deviation (SD) of soluble inflammatory cytokines and chemokines measured in the supernatant of PBMCs either in the absence (italic) or in the presence (bold italic) o treatment with GDNF and GFRα1.

<p>Levels (pg/ml) and standard deviation (SD) of soluble inflammatory cytokines and chemokines measured in the supernatant of PBMCs either in the absence (italic) or in the presence (bold italic) o treatment with GDNF and GFRα1.</p

Phenotypic distribution and levels of expression of RET receptor on circulating immune cell subsets.

<p>(A) Flow cytometric dot plot (upper line) and histogram (lower line) graphs showing a representative example from an healthy donor of CD14^pos monocytes, CD56^pos (NK cells), CD3^pos (T cells) and CD20^pos (B cells) lymphocytes expressing RET receptor (black line) compared to isotype control (gray shaded histograms). (B) Summary graph of dot plots with medians (horizontal black bars) showing the mean fluorescence intensities (MFIs) of RET receptor on immune cells (black symbols) compared to that of isotype controls (open white symbols) from 17 healthy donors.</p