Figure 2
Figure 2. Retrovirus integration sites, genomic rearrangements, and altered gene expressions in mice with B-cell leukemia. (A) Schematic models of retrovirus integration sites. The retrovirus integration sites are indicated by vertical arrows. The left panel illustrates the transgene structure, where the mouse TEC promoter, p210BCR/ABL cDNA, and polyA and splicing signals are shown by dotted, filled, and shaded boxes, respectively. In mice nos. 17 and 30, retroviruses were integrated approximately 1.5-kb and approximately 200-bp upstream of the transcriptional initiation site, respectively. In the right panel, the noncoding and coding regions of Zfp423 exon 1 are shown by blank and filled boxes, respectively. In mice nos. 18 and 23, the viral integration occurred almost in the same site, approximately 100-bp upstream of the translational initiation site. The positions of probes used for Southern blots are also shown. (B) Southern blots to confirm the CISs as major integration sites. Genomic DNAs extracted from the spleen of a control transgenic mouse (C) and tumor tissues of the diseased mice (nos. 17, 30, 18, and 23) were digested with BamHI and blotted with a DNA fragment adjacent to the integration site. Probe A (A) was used for transgene rearrangement (nos. 17 and 30, left panel) and probe B was used for Zfp423 gene rearrangement (nos. 18 and 23, right panel). The positions of germline (G) and rearranged bands are indicated by → and ◀, respectively. Molecular markers are shown on the left. (C) Enhanced expression of p210BCR/ABL in mice nos. 17 and 30 and up-regulated expression of Zfp423 in mice nos. 18 and 23. For detecting p210BCR/ABL message, 20 μg total RNAs extracted from the spleen of a control p210BCR/ABL transgenic mouse (C) and tumor tissues of the diseased mice (nos. 17, 30, 18, and 23) were blotted with p210BCR/ABL cDNA (top left panel) and for detecting Zfp423 message, 3 μg mRNAs from the same tissues were blotted with a part of Zfp423 cDNA (top right panel). The result of β-actin hybridization is shown as an internal control. Molecular markers are shown on the left and the positions of p210BCR/ABL and Zfp423 messages are indicated by →. Enhanced expression of p210BCR/ABL protein in mice nos. 17 and 30 was detected by blotting the proteins extracted from the same tissues with an anti-ABL antibody (bottom left panel). Protein markers are shown on the left and the positions of p210BCR/ABL and c-ABL (145 kDa) are indicated by →.

Retrovirus integration sites, genomic rearrangements, and altered gene expressions in mice with B-cell leukemia. (A) Schematic models of retrovirus integration sites. The retrovirus integration sites are indicated by vertical arrows. The left panel illustrates the transgene structure, where the mouse TEC promoter, p210BCR/ABL cDNA, and polyA and splicing signals are shown by dotted, filled, and shaded boxes, respectively. In mice nos. 17 and 30, retroviruses were integrated approximately 1.5-kb and approximately 200-bp upstream of the transcriptional initiation site, respectively. In the right panel, the noncoding and coding regions of Zfp423 exon 1 are shown by blank and filled boxes, respectively. In mice nos. 18 and 23, the viral integration occurred almost in the same site, approximately 100-bp upstream of the translational initiation site. The positions of probes used for Southern blots are also shown. (B) Southern blots to confirm the CISs as major integration sites. Genomic DNAs extracted from the spleen of a control transgenic mouse (C) and tumor tissues of the diseased mice (nos. 17, 30, 18, and 23) were digested with BamHI and blotted with a DNA fragment adjacent to the integration site. Probe A (A) was used for transgene rearrangement (nos. 17 and 30, left panel) and probe B was used for Zfp423 gene rearrangement (nos. 18 and 23, right panel). The positions of germline (G) and rearranged bands are indicated by → and ◀, respectively. Molecular markers are shown on the left. (C) Enhanced expression of p210BCR/ABL in mice nos. 17 and 30 and up-regulated expression of Zfp423 in mice nos. 18 and 23. For detecting p210BCR/ABL message, 20 μg total RNAs extracted from the spleen of a control p210BCR/ABL transgenic mouse (C) and tumor tissues of the diseased mice (nos. 17, 30, 18, and 23) were blotted with p210BCR/ABL cDNA (top left panel) and for detecting Zfp423 message, 3 μg mRNAs from the same tissues were blotted with a part of Zfp423 cDNA (top right panel). The result of β-actin hybridization is shown as an internal control. Molecular markers are shown on the left and the positions of p210BCR/ABL and Zfp423 messages are indicated by →. Enhanced expression of p210BCR/ABL protein in mice nos. 17 and 30 was detected by blotting the proteins extracted from the same tissues with an anti-ABL antibody (bottom left panel). Protein markers are shown on the left and the positions of p210BCR/ABL and c-ABL (145 kDa) are indicated by →.

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