Figure 1.
Identification of the NUP98-JADE2 fusion. (A) Left, May-Grünwald-Giemsa staining of leukemic blasts in the diagnostic bone marrow (BM). The blasts were medium to large size with fine nuclear chromatin and nucleoli showing hypergranular cytoplasm. Occasional cells with 1 to 2 Auer rods (arrow) were seen but no faggot cell was detected. Right, Sudan Black B-stained positive blasts. Original magnification ×1000. (B) A karyotype performed on the diagnostic BM revealed 46,XX,t(5;11)(q31;p15). (C) Upper, a schematic diagram showing NUP98, JADE2, and NUP98-JADE2 generated by ProteinPaint. The phenylalanine-glycine (FG)/glycine-leucine-phenylalanine-glycine (GLFG) repeats and Gle2-binding-sequence (GLEBS) domain in the amino-terminal portion of NUP98 are retained in the fusion protein. Lower, RT-PCR analysis of NUP98-JADE2 (NJ) and JADE2-NUP98 (JN) fusions in the diagnostic BM. Whole BM cells were used for the analysis. Amplification of GAPDH served as the control. Expected products are indicated by arrowheads. Sanger sequencing of the NJ PCR product is also shown. (D) Immunofluorescence analysis of NUP98-JADE2 and wild-type JADE2. HeLa cells were transfected with pCMV-HA-NUP98-JADE2 or pCMV-HA-JADE2, and the tagged proteins were detected as described in supplemental Methods. DAPI was used for nuclear staining. Original magnification ×1000. (E) Interaction between NUP98-JADE2 and wild-type JADE2. 293T cells were transfected with the indicated Myc- and HA-tagged expression vectors. Myc-tagged proteins were immunoprecipitated and samples were analyzed by immunoblotting for NUP98-JADE2 (NJ) and JADE2 (J2) detection. Similar results were obtained when HA-tagged proteins were immunoprecipitated before immunoblotting (data not shown). (F) Altered subcellular distribution of wild-type JADE2 in the presence of NUP98-JADE2. Upper, HeLa cells were cotransfected with both pCMV-HA-NUP98-JADE2 and pCMV-Myc-JADE2, and the tagged proteins were detected as mentioned previously. Original magnification ×1000. Lower, HeLa cells were transfected with the indicated Myc- and HA-tagged expression vectors. Protein lysates were analyzed by immunoblotting. No apparent change in JADE2 expression was noted when NUP98-JADE2 was coexpressed.

Identification of the NUP98-JADE2 fusion. (A) Left, May-Grünwald-Giemsa staining of leukemic blasts in the diagnostic bone marrow (BM). The blasts were medium to large size with fine nuclear chromatin and nucleoli showing hypergranular cytoplasm. Occasional cells with 1 to 2 Auer rods (arrow) were seen but no faggot cell was detected. Right, Sudan Black B-stained positive blasts. Original magnification ×1000. (B) A karyotype performed on the diagnostic BM revealed 46,XX,t(5;11)(q31;p15). (C) Upper, a schematic diagram showing NUP98, JADE2, and NUP98-JADE2 generated by ProteinPaint. The phenylalanine-glycine (FG)/glycine-leucine-phenylalanine-glycine (GLFG) repeats and Gle2-binding-sequence (GLEBS) domain in the amino-terminal portion of NUP98 are retained in the fusion protein. Lower, RT-PCR analysis of NUP98-JADE2 (NJ) and JADE2-NUP98 (JN) fusions in the diagnostic BM. Whole BM cells were used for the analysis. Amplification of GAPDH served as the control. Expected products are indicated by arrowheads. Sanger sequencing of the NJ PCR product is also shown. (D) Immunofluorescence analysis of NUP98-JADE2 and wild-type JADE2. HeLa cells were transfected with pCMV-HA-NUP98-JADE2 or pCMV-HA-JADE2, and the tagged proteins were detected as described in supplemental Methods. DAPI was used for nuclear staining. Original magnification ×1000. (E) Interaction between NUP98-JADE2 and wild-type JADE2. 293T cells were transfected with the indicated Myc- and HA-tagged expression vectors. Myc-tagged proteins were immunoprecipitated and samples were analyzed by immunoblotting for NUP98-JADE2 (NJ) and JADE2 (J2) detection. Similar results were obtained when HA-tagged proteins were immunoprecipitated before immunoblotting (data not shown). (F) Altered subcellular distribution of wild-type JADE2 in the presence of NUP98-JADE2. Upper, HeLa cells were cotransfected with both pCMV-HA-NUP98-JADE2 and pCMV-Myc-JADE2, and the tagged proteins were detected as mentioned previously. Original magnification ×1000. Lower, HeLa cells were transfected with the indicated Myc- and HA-tagged expression vectors. Protein lysates were analyzed by immunoblotting. No apparent change in JADE2 expression was noted when NUP98-JADE2 was coexpressed.

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