Supplemental material for Rare variants in <i>RNF213</i>, a susceptibility gene for moyamoya disease, are found in patients with pulmonary hypertension and aggravate hypoxia-induced pulmonary hypertension in mice

<p>Supplemental material for Rare variants in <i>RNF213</i>, a susceptibility gene for moyamoya disease, are found in patients with pulmonary hypertension and aggravate hypoxia-induced pulmonary hypertension in mice by Hatasu Kobayashi, Risako Kabata, Hideyuki Kinoshita, Takaaki Morimoto, Koh Ono, Midori Takeda, Jungmi Choi, Hiroko Okuda, Wanyang Liu, Kouji H. Harada, Takeshi Kimura, Shohab Youssefian and Akio Koizumi in Pulmonary Circulation</p

Imaging data of II-2 in Family 1.

<p>(A) MRA image. TOF-3D MRA verifies typical steno-occlusive changes of the circle of Willis. Distal T segments of both internal carotid arteries are occluded and basal moyamoya vessels are clearly seen (anteroposterior view, left panel). Typical “puff-of-smoke” look of moyamoya vessels. Internal carotid arteries are relatively hypoplastic compared with the vertebrobasilar system (lateral view, right panel). (B) Digital subtraction angiography. Catheterization angiography of left vertebral artery (left panel), left carotid artery (middle panel), and right carotid artery (right panel). (C) Transcranial color-coded sonography. Severely dampened flow in the M1 segment of the left middle cerebral artery.</p

MAF in database and prediction of functional change of identified RNF213 variants.

<p>MAF in database and prediction of functional change of identified RNF213 variants.</p

Migration assay using HUVECs transfected with <i>RNF213</i> D4013N, R4019C and V4146A.

<p>Representative images are shown in upper panel. The re-endothelialized areas were quantified by imaging analysis (lower panel). “Vector” represents backbone vector, not including <i>RNF213</i>. Data with bars represent mean ± SD (<i>n</i> = 3 or 4). *<i>P</i> < 0.05 compared with vector, #<i>P</i> < 0.05 compared with WT according to Student’s <i>t</i>-test.</p

Demographic and clinical characteristics of the primary study participants.

<p>Demographic and clinical characteristics of the primary study participants.</p

Identification of <i>RNF213</i> rare variants in three families.

<p>(A) Pedigree chart and genotypes of <i>RNF213</i> rare variants and microsatellite markers of the three families. Filled and unfilled symbols indicate affected and unaffected individuals, respectively. Squares and circles represent males and females, respectively. Arrows indicate index case. (B) Sequence chromatography of the identified <i>RNF213</i> rare variants. (C) Haplotype for p.R4019C and p.E4042K determined by cloning in II-1 in Family 2.</p

Demographic and clinical characteristics of the replication study participants.

<p>Demographic and clinical characteristics of the replication study participants.</p

Clinical characteristics of the primary study population according to the <i>RNF213</i> p.R4810K genotype (GG vs. GA+AA).

<p>Clinical characteristics of the primary study population according to the <i>RNF213</i> p.R4810K genotype (GG vs. GA+AA).</p

Multivariate analysis of the risk of coronary artery disease among the primary study population without a history of diabetes mellitus.

<p>Multivariate analysis of the risk of coronary artery disease among the primary study population without a history of diabetes mellitus.</p

Homology of identified RNF213 variants.

<p>Homology of identified RNF213 variants.</p