1. The (4;11)(q21;p15) Translocation Fuses the NUP98 andRAP1GDS1 Genes and Is Recurrent in T-Cell Acute Lymphocytic Leukemia
by Damian J. Hussey, Mario Nicola, Sarah Moore, Gregory B. Peters, and Alexander Dobrovic
Blood
Volume 94(6):
September 15, 1999
©1999 by American Society of Hematology

2. Position of the chromosome 11 breakpoint with respect to the 11p15
Position of the chromosome 11 breakpoint with respect to the 11p15.5 markers used for FISH and PCR mapping.
Position of the chromosome 11 breakpoint with respect to the 11p15.5 markers used for FISH and PCR mapping. NUP98 lies within the candidate breakpoint region indicated by the arrowed line. The β chain of hemoglobin (HBBC) and the H-ras oncogene (HRAS) are at the extremities of 11p15.5. C and T denote centromeric and telomeric, respectively.
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology

3. PCR analysis of the der(4) and der(11) containing somatic cell hybrids
PCR analysis of the der(4) and der(11) containing somatic cell hybrids. m is the pUC19/Hpa II molecular weight marker, H is normal human, M is mouse, 4 is the der(4) hybrid, and 11 is the der(11) hybrid.
PCR analysis of the der(4) and der(11) containing somatic cell hybrids. m is the pUC19/Hpa II molecular weight marker, H is normal human, M is mouse, 4 is the der(4) hybrid, and 11 is the der(11) hybrid. (A) NUP98 exon B PCR product digested withTaq I. Mouse and human NUP98 cDNA sequences are highly conserved and the exon B PCR also amplified mouse NUP98. The mouse and human exon B PCR products were distinguished by a TaqI restriction site, which is present in the mouse product but absent in the human product. (B) NUP98 exon C PCR product.
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology

4. Nucleotide and amino acid sequences around the junctions of the (A) NRG and (B) NRG2 fusion transcripts (Genbank accession nos.AF and AF133333, respectively).
Nucleotide and amino acid sequences around the junctions of the (A) NRG and (B) NRG2 fusion transcripts (Genbank accession nos.AF and AF133333, respectively).
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology

5. RT-PCR analysis of NRG and RGN fusion transcripts in 3 t(4;11)(q21;p15) patients.
RT-PCR analysis of NRG and RGN fusion transcripts in 3 t(4;11)(q21;p15) patients. P1, P2, and P3 are RT-PCR products from peripheral blood mononuclear cells from the patients. C1 and C2 are RT-PCR products from peripheral blood mononuclear cells of normal donors. Samples marked with a minus sign are negative control RT-PCRs without reverse transcriptase. H2O controls are negative control RT-PCRs without target. The lane marked m contains both SPP1/EcoRI and pUC19/Hpa II molecular weight markers. The most prominent bands in the RGN PCR of patient no. 3 are alternative splicings of RGN with and without exon B.
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology

6. Northern analysis of NRG expression.
Northern analysis of NRG expression. (A) Hybridization using a NUP98 cDNA probe. (B) Hybridization of the same membrane with a RAP1GDS1 cDNA probe. (C) 18S rRNA from the ethidium bromide-stained gel before transfer. RNA was isolated from 2 normal controls (C1 and C2) and from the 3 patients (P1, P2, and P3). pres is a presentation sample, rem is a remission sample, and rel is a relapse sample. Each lane contains 5 μg of total RNA from peripheral blood mononuclear cells, except that P1 rem contains 5 μg of total RNA from bone marrow. The lane marked m is a RNA ladder (Promega). The band in this lane in (C) is marker and not 18S RNA. N indicates theNUP and 7.25-kb bands. The 7.25-kb band is a precursor that also contains the NUP96 coding sequence.47 RG indicates the 2.8- and 4.1-kb RAP1GDS1 bands. NRG indicates the 4.4-kb NRG transcript. NRG2 in patient no. 3 is not indicated, because it is not distinguishable from the 4.05-kbNUP98 and 4.1-kb RAP1GDS1 bands. The arrowheads indicate higher molecular weight transcripts that hybridize with both the NUP98 and RAP1GDS1 probes.
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology

7. Multiple tissue Northern analysis of RAP1GDS1.
Multiple tissue Northern analysis of RAP1GDS1. Each lane contains 2 μg of polyA RNA. (A) Hybridization with aRAP1GDS1 cDNA probe shows two predominant bands of 4.1 and 2.8 kb. (B) Hybridization with a β-actin cDNA probe (Clontech).
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology

8. Schematic representation of the NUP98, smgGDS, NRG, and NRG2 proteins.
Schematic representation of the NUP98, smgGDS, NRG, and NRG2 proteins. Vertical arrowheads represent breakpoints in NUP98 and smgGDS. FG, FG (phenylalanine-glycine) repeat-rich areas; GLEBS, GLEBS (Gle2p-binding motif) -like motif48; NRM, nucleoporin RNA binding motif; ARMADILLO, tandem armadillo repeats.
Damian J. Hussey et al. Blood 1999;94:
©1999 by American Society of Hematology