PMC5693504_01
Patient's metaphase spread, with the arrow indicating the marker chromosome
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Fig1
Although DAPI staining showed that this chromosome looks such as a small acrocentric chromosome, localization of the telomeric probe revealed no signals on it (Fig. 1).
Examples of fluorescent in situ hybridization: a Localization of human chromosome X painting probe (red) on chromosomes of the patient. b Localization of microdissected probe of marker chromosome on chromosomes of the patient. c Localization of microdissected probe of marker chromosome on chromosomes of the healthy control after G-banding (d). Box on (c) is localization of signal on G-banded chromosome X. Arrows indicate signal localization
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Fig2
FISH of the set of human whole chromosomes-specific probes showed the localization of chromosome X painting probe on the whole length of the marker chromosome (Fig. 2a). In the patient cells each probe painted the marker chromosome and the centromeric region of chromosome X (Fig. 2b). In the healthy control, probes painted the centromeric region of chromosome X only (Fig. 2c, d). This localization showed the marker chromosome was derived from region Xp11.23-q13.2. (Fig. 2c - box).  
Results of PCR screening of marker chromosomes for XIST gene presence. Primers: CH3R/F on lines 1-5; CHXR/F on lines 6-8, 11, 12; XISTF/R on lines 13-17. DNA templates: chromosome 3 painting probe on lines 1, 6, 13; chromosome X painting probe on lines 2, 7, 14; microdissection-derived marker chromosome (mar1) on lines 3, 8, 15; microdissection-derived marker chromosome (mar2) on lines 4, 11, 16; negative controls on lines 5, 12, 17; 100 bp ladder on lines 9, 10, 18
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Fig3
Both analyzed marker chromosomes do not contain XIST sequences whereas X centromeric sequences are presented in marker chromosomes painting probes (Fig. 3).