American Society of Hematology

Fig. 2.

$Fig. 2. Effect of overexpression of GLO1 on antitumor agent-induced apoptosis. / pFLAG (mock) or pFLAG-GLO1 (GLO1) expression plasmids were transiently cotransfected into Jurkat cells with pHook-1 as described in “Materials and methods,” and the transfected cells were selected using magnetic beads coated with phOx. (A) The expression of FLAG-tagged GLO1 in Jurkat cells was examined by anti-FLAG Western blot analysis. (B) Cytosolic fractions were isolated, and GLO1 activity was performed as described in “Materials and methods.” The means ± SD of triplicate cultures are shown. (C) Cells were treated with various concentrations of ADM or VP-16 for 24 hours, and cytotoxicity was determined by MTS method. The means ± SD of triplicate cultures are shown. (D) Cells were treated with 1.2 μmol/L of ADM for 14 hours, and caspase-3–like protease activity was measured with the specific fluorgenic substrate Ac-DEVD-MCA. Results are the means ± SD in triplicate. (E) Cells were treated with 0.5 μmol/L of ADM for 24 hours and were stained with Hechest 33342.$

Effect of overexpression of GLO1 on antitumor agent-induced apoptosis.

pFLAG (mock) or pFLAG-GLO1 (GLO1) expression plasmids were transiently cotransfected into Jurkat cells with pHook-1 as described in “Materials and methods,” and the transfected cells were selected using magnetic beads coated with phOx. (A) The expression of FLAG-tagged GLO1 in Jurkat cells was examined by anti-FLAG Western blot analysis. (B) Cytosolic fractions were isolated, and GLO1 activity was performed as described in “Materials and methods.” The means ± SD of triplicate cultures are shown. (C) Cells were treated with various concentrations of ADM or VP-16 for 24 hours, and cytotoxicity was determined by MTS method. The means ± SD of triplicate cultures are shown. (D) Cells were treated with 1.2 μmol/L of ADM for 14 hours, and caspase-3–like protease activity was measured with the specific fluorgenic substrate Ac-DEVD-MCA. Results are the means ± SD in triplicate. (E) Cells were treated with 0.5 μmol/L of ADM for 24 hours and were stained with Hechest 33342.

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