Figure 2
Figure 2. JQ1 treatment of murine T-ALL cells results in cell-cycle arrest followed by apoptosis. Murine T-ALL cell lines were treated with JQ1 (250 nM) or with the GSI (Compound E, 1 μM), unless otherwise noted. (A) JQ1-induced cell-cycle arrest is evident at 24 hours. Murine T-ALL cell lines were left untreated or treated with JQ1 or Compound E for 24 hours and then stained with propidium iodide followed by flow cytometry. Four mouse T-ALL cell lines were analyzed; 1 representative plot from mouse T-ALL 5109 is shown. (B) JQ1 treatment induces cell-cycle arrest followed by apoptosis. Murine T-ALL cell line 5109 was treated with JQ1 or Compound E for 24, 48, and 96 hours and the percentage of cells in the subG1, G0/G1, and S phases determined. (C) Mouse T-ALL cell lines are more sensitive to treatment with JQ1 than GSI. Four mouse T-ALL cell lines (4673, 5109, 5059, and 6124) were treated with DMSO, JQ1, or the GSI (Compound E) for 96 hours and the total number of viable cells was calculated via trypan blue exclusion assay. The results are averages of 3 independent experiments and error bars represent standard error of the mean (SEM). P < .0001 for all JQ1-treated cell lines, *P < .05, **P < .01, ***P < .001. (D) JQ1 induces apoptosis of mouse T-ALL cells. Four mouse T-ALL cell lines were treated with vehicle, JQ1, or Compound E and the percentage of apoptotic cells determined by Annexin V and 7AAD staining followed by flow cytometry. The results are averages of 3 independent experiments and error bars represent SEM. *P < .05, **P < .01, ***P < .001. (E) Reduced c-Myc mRNA levels in JQ1- and GSI-treated mouse T-ALL cells. RNA was isolated from leukemic cultures treated with vehicle, JQ1, or Compound E for 48 hours and c-Myc expression was analyzed by quantitative real-time PCR. Copy number was normalized to β-actin using the ΔΔCT method. The results are averages of 3 independent experiments and error bars represent SEM. P < .0001 for all JQ1-treated cell lines *P < .05, **P < .01, ***P < .001. (F) c-Myc protein levels were reduced upon treatment with JQ1. Protein was isolated from mouse T-ALL cells treated with DMSO, JQ1, or GSI (Compound E) for 48 hours and c-Myc and β-actin protein levels determined by immunoblotting.

JQ1 treatment of murine T-ALL cells results in cell-cycle arrest followed by apoptosis. Murine T-ALL cell lines were treated with JQ1 (250 nM) or with the GSI (Compound E, 1 μM), unless otherwise noted. (A) JQ1-induced cell-cycle arrest is evident at 24 hours. Murine T-ALL cell lines were left untreated or treated with JQ1 or Compound E for 24 hours and then stained with propidium iodide followed by flow cytometry. Four mouse T-ALL cell lines were analyzed; 1 representative plot from mouse T-ALL 5109 is shown. (B) JQ1 treatment induces cell-cycle arrest followed by apoptosis. Murine T-ALL cell line 5109 was treated with JQ1 or Compound E for 24, 48, and 96 hours and the percentage of cells in the subG1, G0/G1, and S phases determined. (C) Mouse T-ALL cell lines are more sensitive to treatment with JQ1 than GSI. Four mouse T-ALL cell lines (4673, 5109, 5059, and 6124) were treated with DMSO, JQ1, or the GSI (Compound E) for 96 hours and the total number of viable cells was calculated via trypan blue exclusion assay. The results are averages of 3 independent experiments and error bars represent standard error of the mean (SEM). P < .0001 for all JQ1-treated cell lines, *P < .05, **P < .01, ***P < .001. (D) JQ1 induces apoptosis of mouse T-ALL cells. Four mouse T-ALL cell lines were treated with vehicle, JQ1, or Compound E and the percentage of apoptotic cells determined by Annexin V and 7AAD staining followed by flow cytometry. The results are averages of 3 independent experiments and error bars represent SEM. *P < .05, **P < .01, ***P < .001. (E) Reduced c-Myc mRNA levels in JQ1- and GSI-treated mouse T-ALL cells. RNA was isolated from leukemic cultures treated with vehicle, JQ1, or Compound E for 48 hours and c-Myc expression was analyzed by quantitative real-time PCR. Copy number was normalized to β-actin using the ΔΔCT method. The results are averages of 3 independent experiments and error bars represent SEM. P < .0001 for all JQ1-treated cell lines *P < .05, **P < .01, ***P < .001. (F) c-Myc protein levels were reduced upon treatment with JQ1. Protein was isolated from mouse T-ALL cells treated with DMSO, JQ1, or GSI (Compound E) for 48 hours and c-Myc and β-actin protein levels determined by immunoblotting.

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