PMC4743364_01
Montages of NIRFLI of the lower extremities of (a) the proband and (b) her mother. While the mother had well-defined lymphatics, she also had areas with dermal backflow as shown in the inset images of (b)
13221_2016_35_Fig1_HTML.jpg
PMC4743364_01_13221_2016_35_Fig1_HTML.jpg
Fig1
As shown by NIRFLI in Fig. 1b, the affected daughter had extensive areas of dermal backflow in her lower legs, as commonly observed in LE subjects, but possessed a unique, uncommon flow pattern of lymph movement that appeared in "sheets" rather than within distinct vessel lumens (Additional file 1: Video 1). While the proband's mother primarily had well-defined and linear lymphatics (Fig. 1b) as typically seen in healthy subjects, she nonetheless presented with smaller areas of dermal backflow and tortuous lymphatics as shown in the insets of Fig. 1b. TGAGGTTGGGAGCCGGTAGAGG and the reverse direction using primer GTACCGACAGGGTATGTGAAGGCG (see Additional file 3: Figure S1). The segregation results were perfect, the variant was present in all the affected and was not present in any of the unaffected.
Pedigree of the family showing members diagnosed with lymphedema (filled black), unknown phenotype (filled gray) and unaffected (open) subjects. Arrow indicates the proband. The small 's' at the left top corner of the subjects indicates when the subject's DNA was sequenced. A diagonal line indicates that the patient is deceased
13221_2016_35_Fig2_HTML.jpg
PMC4743364_01_13221_2016_35_Fig2_HTML.jpg
Fig2
The pedigree chart in Fig. 2 shows that the diagnosed disorder segregates in an autosomal dominant manner.
a Image from the Integrative genomics viewer, showing a 40 bp genomic region containing the variant. Each one of the samples is loaded into the panel. Each panel shows two windows, the top window displays the number of reads, and the allele fraction and the lower window show the alignments of the experimental reads. The bottom panel displays the transcripts with the open reading frames. b A cartoon representing the predicted protein structure of CELSR1 (uniprot id Q9NYQ6). An arrow represents the location of the mutation. The cartoon was inspired by the publication of Lei, et al., 2014
13221_2016_35_Fig3_HTML.jpg
PMC4743364_01_13221_2016_35_Fig3_HTML.jpg
Fig3
In summary, the ultra-rare damaging variant was present in five affected members and not present in six unaffected family members (Fig. 3a).