PMC4962816_01
Sequences of junctional regions of three different IKZF1 deletions in BM cells genomic DNA of the patient. Positions of primers used for RQ-PCR are underlined by a solid line, probes - the dotted line. Intron sequences before and after the deletion breakpoint colored in green and blue, non-matrix nucleotides - in red. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
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At the relapse another one Ex3-6r deletion was found (Fig. 1).
iFISH analysis of IKZF1 gene, objectives at 100x magnification. Purple fluorescence signals marked IKZF1 locus, the nuclei stained with DAPI II (blue). One separate purple signal indicates lack of IKZF1 (intragenic deletion). (1-3) Diagnosis; (4-6) Relapse; (7-9) Control BM cells from patient with IKZF1 wild type. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
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IKZF1 alleles, 2.5% with one, and 2.5% with three signals, demonstrating a method error level of 5%. 70% of patient's BM cells at diagnosis showed deletion of one IKZF1 allele and 26% cells brought both normal alleles. In the BM at relapse 59% of cells were heterozygous for the IKZF1 deletions and 33% of cells with normal alleles were observed (Fig. 2).
Confocal microscopy was used to examine the localization of wild-type and mutant Ikaros proteins. Nuclei stained with propidium iodine (PI, red), and Ikaros proteins were visualized by PE Mouse Anti-Human Ikaros antibodies (green), objectives at 100x magnification. Micron scale 10 microm. (1-3) Abnormal cytoplasmic compartmentalization patterns of Ikaros protein (green) in patient's BM cells at diagnosis; (4-6) Nuclear localization of Ikaros protein (green) in control ALL BM cells. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
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