Genetic mechanisms leading to primary amenorrhea in balanced X-autosome translocations  Mariana Moysés-Oliveira, M.Sc., Roberta dos Santos Guilherme, M.Sc., Anelisa Gollo Dantas, B.S., Renata Ueta, B.S., Ana Beatriz Perez, Ph.D., Mauro Haidar, Ph.D., Rosane Canonaco, M.D., Vera Ayres Meloni, M.D., Nadezda Kosyakova, Ph.D., Thomas Liehr, Ph.D., Gianna Maria Carvalheira, Ph.D., Maria Isabel Melaragno, Ph.D.  Fertility and Sterility  Volume 103, Issue 5, Pages 1289-1296.e2 (May 2015) DOI: 10.1016/j.fertnstert.2015.01.030 Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Breakpoint definition for patient 1. (A) Partial G-banding karyotype and ideogram of the chromosomes involved in the rearrangement. (B, C) Array painting results showing (B) X-chromosomal breakpoint and (C) chromosome 3 breakpoint in the color change region. (D, E) FISH in metaphase cells using (D) RP11-34P8 and (E) RP11-1072D12 probes, showing three hybridization signals, including split signals in the derivative chromosomes. Fertility and Sterility 2015 103, 1289-1296.e2DOI: (10.1016/j.fertnstert.2015.01.030) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Region affected by X-chromosome breakpoints in patients 1 and 2. (A) Partial G-banding karyotype and ideogram of the chromosomes involved in patient 2's rearrangement. (B) Genome view of the Xq13.3 cytoband and the FGF16 syntenic block, including breakpoints in patients 1 and 2 (BP, red bars), the microduplication (blue bar) described by Dudding et al. (35), and the genes conserved in the syntenic block in red (ZDHHC15 - FGF16 - ATRX). The FGF16 gene is located approximately 2 Mb downstream of patient 2's breakpoint. Fertility and Sterility 2015 103, 1289-1296.e2DOI: (10.1016/j.fertnstert.2015.01.030) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Region affected by autosomal breakpoint in patient 3. (A) Partial G-banding karyotype and ideogram of the chromosomes involved in the rearrangement. (B) Genome view of the region downstream to chromosome 2 breakpoint in patient 3, including segment affected by the autosomal breakpoint of patient 3 (BP, red bar) and the FHL2 gene, located 121.6 kb downstream of the breakpoint. Fertility and Sterility 2015 103, 1289-1296.e2DOI: (10.1016/j.fertnstert.2015.01.030) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Chromosome 9 breakpoint definition by FISH for patient 2 using centromeric and pericentromeric probes for chromosome 9. (A) Scheme of the regions assessed by the centromeric and pericentromeric probes that were used, and the breakpoint localization indicated by the black line. (B) FISH signals in the normal chromosome 9, derivative chromosome 9, and derivative chromosome X; regions investigated by the probes are indicated on top, and intensities of the signals are represented on right. The probe for chromosome 9 centromere (labeled in green) showed three signals: one in normal chromosome 9 and one in each derivative chromosome, indicating that the autosomal breakpoint affected the chromosome 9 centromeric region. (C) FISH in metaphase cells using centromeric probes for chromosomes X and 9, labeled in green and red, respectively, showing that the autosomal breakpoint appeared in the region covered by the chromosome 9 centromeric probe. Fertility and Sterility 2015 103, 1289-1296.e2DOI: (10.1016/j.fertnstert.2015.01.030) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions