Figure 6.
Combination of BCL6 inhibitor and AraC mediates synergistic effects. (A) Five primary AML specimens (AML54, AML74, AML95, AML98, and AML109) were subjected to ex vivo combination therapy using AraC and FX1 with various combinations of limiting dilution assay. Matrices indicate calculated synergy scores obtained from each pair of combination doses. Synergy scores were calculated by using SynergyFinder software. The highest single agent (HSA) algorithm was used to calculate synergy scores, and color keys indicate the scores in heatmaps. (B) Colony formation assay in primary AML cells treated with AraC, BPI, or combination of both. Statistical significance between each of the conditions was calculated by one-way analysis of variance. (C) Evaluation of cell deaths in primary AML cells (AML33) treated with AraC in the presence or absence of siRNA-induced BCL6 knockdown. Primary AML cells (AML33) were transiently transfected with control or BCL6 siRNA; the cells were then treated with AraC in duplicates. Relative percent dead cells were calculated by normalization to untreated cells. Comparing the relative percent dead cells vs that of parental AML33 cells treated with AraC, the increase of dead cells with each siRNA is presented as relative values. Averages were obtained from duplicates, and error bars represent standard deviations. (D) The same series of primary AML33 cells as (C) were subjected to colony formation assays after 24 hours of ex vivo treatment with AraC. The number of colonies was evaluated after 14 days. Colony counts were compared between untreated and AraC treatment to obtain relative colony numbers, and relative colony number reductions were obtained by comparing the relative values vs parental AML cells. In (C) and (D), statistical significance was evaluated by using an unpaired Student t test. (E) Representative photographs of colony formation assays. Thirty-five (7 × 5) pictures of single fields were tiled to show entire views of each plate. (F) Representative fields marked by the box in (E) were presented in zoomed views. CFU, colony-forming unit; NS, not significant; NTC, nontargeting control.

Combination of BCL6 inhibitor and AraC mediates synergistic effects. (A) Five primary AML specimens (AML54, AML74, AML95, AML98, and AML109) were subjected to ex vivo combination therapy using AraC and FX1 with various combinations of limiting dilution assay. Matrices indicate calculated synergy scores obtained from each pair of combination doses. Synergy scores were calculated by using SynergyFinder software. The highest single agent (HSA) algorithm was used to calculate synergy scores, and color keys indicate the scores in heatmaps. (B) Colony formation assay in primary AML cells treated with AraC, BPI, or combination of both. Statistical significance between each of the conditions was calculated by one-way analysis of variance. (C) Evaluation of cell deaths in primary AML cells (AML33) treated with AraC in the presence or absence of siRNA-induced BCL6 knockdown. Primary AML cells (AML33) were transiently transfected with control or BCL6 siRNA; the cells were then treated with AraC in duplicates. Relative percent dead cells were calculated by normalization to untreated cells. Comparing the relative percent dead cells vs that of parental AML33 cells treated with AraC, the increase of dead cells with each siRNA is presented as relative values. Averages were obtained from duplicates, and error bars represent standard deviations. (D) The same series of primary AML33 cells as (C) were subjected to colony formation assays after 24 hours of ex vivo treatment with AraC. The number of colonies was evaluated after 14 days. Colony counts were compared between untreated and AraC treatment to obtain relative colony numbers, and relative colony number reductions were obtained by comparing the relative values vs parental AML cells. In (C) and (D), statistical significance was evaluated by using an unpaired Student t test. (E) Representative photographs of colony formation assays. Thirty-five (7 × 5) pictures of single fields were tiled to show entire views of each plate. (F) Representative fields marked by the box in (E) were presented in zoomed views. CFU, colony-forming unit; NS, not significant; NTC, nontargeting control.

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