American Society of Hematology

Fig. 3.

$Fig. 3. Loss of PTEN expression due to genomic DNA rearrangement in OPM2 and Δ47 human MM lines. / (A) Equivalent amounts of cell lysates were separated by SDS-PAGE and immunoblotted with anti-PTEN. (B) Genomic DNA was isolated from the cell lines shown in the figure. 10 μg of genomic DNA was digested with either HindIII or EcoRI overnight and separated by 0.7% agarose gel. DNA transferred membrane was hybridized with 1.2-kb human PTEN cDNA probe. Washed membrane was autoradiographed. DNA size markers are shown. (C) Twenty μg of total RNA from each sample was fractionated using the Trizol Reagent and hybridized with human PTEN cDNA probe (upper left panel). The 3.0- and 5.5-kb PTEN transcripts are marked by arrows. The same membrane was stripped and rehybridized with β-actin probe (lower left panel). After normalization for RNA loading by β-actin control, the RNA levels are arbitrarily plotted by using 7.2 and Δ47 as basal levels for mouse and human lines, respectively (right panel).$

Loss of PTEN expression due to genomic DNA rearrangement in OPM2 and Δ47 human MM lines.

(A) Equivalent amounts of cell lysates were separated by SDS-PAGE and immunoblotted with anti-PTEN. (B) Genomic DNA was isolated from the cell lines shown in the figure. 10 μg of genomic DNA was digested with either HindIII or EcoRI overnight and separated by 0.7% agarose gel. DNA transferred membrane was hybridized with 1.2-kb human PTEN cDNA probe. Washed membrane was autoradiographed. DNA size markers are shown. (C) Twenty μg of total RNA from each sample was fractionated using the Trizol Reagent and hybridized with human PTEN cDNA probe (upper left panel). The 3.0- and 5.5-kb PTEN transcripts are marked by arrows. The same membrane was stripped and rehybridized with β-actin probe (lower left panel). After normalization for RNA loading by β-actin control, the RNA levels are arbitrarily plotted by using 7.2 and Δ47 as basal levels for mouse and human lines, respectively (right panel).

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