Deformability and fragility studies of RBCs.

<p>(A) RBC deformability and relaxation capacity was studied with a microfluidic device [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125580#pone.0125580.ref012" target="_blank">12</a>]. Scale bar 10 μm. (B) Deformability and fragility were also measured with a spleen-mimicking device [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125580#pone.0125580.ref013" target="_blank">13</a>]. The cells of interest were stained with CFSE, mixed with RBCs of a control donor, and passed through a bed of metal beads [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125580#pone.0125580.ref013" target="_blank">13</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125580#pone.0125580.ref014" target="_blank">14</a>]. Retention was measured by the number of cells that did not pass through the bead layer. The percentages of CFSE-positive cells in the retained and downstream sample were compared to that in the upstream sample (set at 5%, depicted as a dotted line in panel B).</p

Pedigree and RBC morphology of the subjects in this study.

<p>(A) Patients who are clinically diagnosed with PKAN are indicated in black, the healthy relatives are indicated in white. (B) Representative pictures of blood films used for classification of cell shape. (C) Cells are classified in discocyte, echinocyte, acanthocyte or otherwise misshapen. The graph depicts the percentages of different cell morphologies in PKAN patients (marked with an asterisk) and their unaffected family members, compared to healthy donors (control 1+2) (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125580#pone.0125580.s001" target="_blank">S1 Fig</a> for other PKAN families).</p

Measurements of fetal haemoglobin (HbF) in red cells of PKAN patients and their relatives.

<p>Black, patients; grey, controls (Ctrls) and family members. The control value is the mean of 6 control samples. The error bar depicts the standard deviation.</p

Microparticle formation by PKAN RBCs as characterized by flow cytometry.

<p>(A) The amount of RBC MPs in plasma. (B) Membrane composition of in vitro generated MPs. Band 3 and glycophorin A (GpA) was measured on MPs collected after 4 and 24h incubation of the RBCs in Ringer solution at 37 °C.</p