PMC6614923_01
PMC6614923_02
PMC6614923_03
Pedigree of the family and the characterization of the deletion breakpoints by gap-PCR. (A) Pedigree of this family. The arrow refers to the proband. The solid square (male) and circle (female) represent the two affected siblings; (B) Gap-PCR of family members using the peripheral blood-derived genomic DNA and primer sets (4334-F/R). Lanes 1-7: Marker, II2, II1, III1, III2, II3, and I1. The expected SATB2 wild allele band of ~4.3 kb is seen in members of II2, II1, III1, III2, II3, and I1. A lower band, consistent with the size of mutant allele, is seen in II1, III1, and III2 but not in II2 or I1. Notice that the father (II1) has a much less intense band of about 1.3 kb in size, which is the same size as his two children (III1 and III2). (C) Sequencing results of the ~4.3-kb band and 1.3-kb band. The intragenic deletion border is chr2: 200,191,313-200,194,324 (hg19), which includes the entire exon 9 and its franking intronic sequences of the SATB2 gene. Blue arrows refer to gap-PCR primer set (4334-F/R) used to detect the breakpoints of the 3kb deletion.
fgene-10-00630-g001.jpg
PMC6614923_03_fgene-10-00630-g001.jpg
f1
They have a 4.9-kb deletion in the 2q33.1 region of chromosome 2 (Figure S1). Peripheral blood samples were collected from the family members including the husband's father (I1), the couple (II1 and II2), and their two children bearing SAS (III1 and III2) ( Figure 1A ).
Quantitative PCR (qPCR) of family members using the peripheral blood-derived genomic DNA. (A) Schematic diagram of location of qPCR primers. Blue arrows refer to qPCR primers; (B) Comparison of gene dosage ratios for the family members using the SATB2-exon 9 primers; (C) Comparison of gene dosage ratios for the family members using the SATB2-intron 8 primers; (D) Comparison of gene dosage ratios for the family members using the SATB2-intron 9 primers. F, normal female; C, control (normal male).
fgene-10-00630-g002.jpg
PMC6614923_03_fgene-10-00630-g002.jpg
f2
Three pairs of primers ( Figure 2A ) were designed to detect potential deletions in this family, including two primer sets outside the deletion region and one primer set inside the deletion.