Figure 3
(A) Family Pedigree Proband 4. Emm RBC phenotypes shown include the North African Proband, who was pregnant with anti-Emm in the plasma, and her parents and 7 siblings. (B) Amplification of PIGG Exons 1 through 13. Amplified products covering the 13 Exons are shown. Arrow indicates the location of the Exon 3 product in the control sample. Exon 3 demonstrates ~ 100-bp deletion in the sample from Proband 4 compared to control. (C) Proband 4 deletion breakpoints. Amplification of Exon 3 in the sample from North African Proband 4 indicated an approximate insertion deletion (indel) in PIGG compared to the control (left). Targeted NGS indicated a breakpoint spanning the region of Intron 2 and Exon 3 compared (top). Analysis showed split reads containing the breakpoint sequencing spanning the deletion in Intron 2 and Exon 3 with 5 extra bases inserted near exon 3 (colored rectangles in alignment), c.361-51_383delinsGACTT, p.(Ala121_Pro128delinsAspPhe). Breakpoint was confirmed by Sanger sequencing (middle, note Sanger traces are reverse complemented since sequencing done using reverse primer). Deletion breakpoint sequence compared to wild type (bottom).
