Fig. 7.
Fig. 7. Overexpression of RHAMMFL and RHAMM−147 in MM, B-lymphoma, and B-CLL, as compared with normal B cells and chronically activated B cells from patients with Crohn’s disease. (A) RHAMMFL and RHAMM−147 were amplified by RT-PCR of 1μg RNA from the BM plasma cells of 6 myeloma patients, from the B cells of 5 myeloma patients, and from the B cells of 5 normal individuals. The same quantity of RNA (1 μg) and cDNA (5 μL) was used in the RT-PCR step for all cell populations analyzed so that the level of RHAMM expression could be compared between the different cell populations in the figure. The quality of RNA isolated from all cell populations was comparable as determined by amplification of histone transcripts from a separate aliquot of the same cDNA (1 μL) from which RHAMM was amplified (data not shown). Markers are 1,114, 900, and 692 bp. A 989-bp fragment was amplified for RHAMMFL and a 842-bp fragment for RHAMM−147. The amplification products were transferred to a nylon membrane and hybridized to a DIG-labeled RHAMM probe. BM plasma cell RHAMM was visualized by colorimetric detection; color development was for 1 hour. MM B-cell RHAMM and normal B cell RHAMM was detected by chemiluminescence; exposure was for 18 hours. (B) RHAMMFLand RHAMM−147 were amplified by RT-PCR of 500 ng RNA from the malignant lymph nodes of 3 patients (m, n, o), the PBMC of 3 CLL patients (p, q, r), the B cells of 3 normal individuals (s, t, u), and the B cells from a patient with Crohn’s disease (v). The same quantity of RNA (500 ng) and cDNA (5 μL) was used in the RT-PCR step for all cell populations analyzed so that the level of RHAMM expression could be compared between the different cell populations in the figure. The quality of RNA isolated from all B-cell populations was comparable as determined by amplification of CD19 transcripts from a separate aliquot of the same cDNA (5 μL) from which RHAMM was amplified (data not shown). A 989-bp fragment was amplified for RHAMMFL and a 842-bp fragment for RHAMM−147; arrows indicate their position. The amplification products were transferred to a nylon membrane and hybridized to a DIG-labeled RHAMM probe. RHAMM was visualized by colorimetric detection; color development was for 1 hour. Markers are 1,114 and 900 bp.

Overexpression of RHAMMFL and RHAMM−147 in MM, B-lymphoma, and B-CLL, as compared with normal B cells and chronically activated B cells from patients with Crohn’s disease. (A) RHAMMFL and RHAMM−147 were amplified by RT-PCR of 1μg RNA from the BM plasma cells of 6 myeloma patients, from the B cells of 5 myeloma patients, and from the B cells of 5 normal individuals. The same quantity of RNA (1 μg) and cDNA (5 μL) was used in the RT-PCR step for all cell populations analyzed so that the level of RHAMM expression could be compared between the different cell populations in the figure. The quality of RNA isolated from all cell populations was comparable as determined by amplification of histone transcripts from a separate aliquot of the same cDNA (1 μL) from which RHAMM was amplified (data not shown). Markers are 1,114, 900, and 692 bp. A 989-bp fragment was amplified for RHAMMFL and a 842-bp fragment for RHAMM−147. The amplification products were transferred to a nylon membrane and hybridized to a DIG-labeled RHAMM probe. BM plasma cell RHAMM was visualized by colorimetric detection; color development was for 1 hour. MM B-cell RHAMM and normal B cell RHAMM was detected by chemiluminescence; exposure was for 18 hours. (B) RHAMMFLand RHAMM−147 were amplified by RT-PCR of 500 ng RNA from the malignant lymph nodes of 3 patients (m, n, o), the PBMC of 3 CLL patients (p, q, r), the B cells of 3 normal individuals (s, t, u), and the B cells from a patient with Crohn’s disease (v). The same quantity of RNA (500 ng) and cDNA (5 μL) was used in the RT-PCR step for all cell populations analyzed so that the level of RHAMM expression could be compared between the different cell populations in the figure. The quality of RNA isolated from all B-cell populations was comparable as determined by amplification of CD19 transcripts from a separate aliquot of the same cDNA (5 μL) from which RHAMM was amplified (data not shown). A 989-bp fragment was amplified for RHAMMFL and a 842-bp fragment for RHAMM−147; arrows indicate their position. The amplification products were transferred to a nylon membrane and hybridized to a DIG-labeled RHAMM probe. RHAMM was visualized by colorimetric detection; color development was for 1 hour. Markers are 1,114 and 900 bp.

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