Figure 2
Characterization of UPN 476081. Cells from 476081 were grown on stroma for 4 days, followed by a 3-day treatment with cytarabine (AraC) or decitabine (DAC). Drug was administered daily. After 3 days of treatment, the number of viable cells (A), annexin-positive cells (B), and total 5-methycytosine content by LC-MS/MS (C) was determined. (D-E) cell cycle profiles were determined after overnight EdU incorporation and DNA content was measured by FxCycle Violet DNA dye. Shown are representative flow plots (D) and distributions of cell cycle phase after drug treatment (E). All experiments were performed in triplicate; error bars represent 95% confidence intervals; asterisk indicates a significant change from mock treatment. (F) CD45dim/SSlow blast gate (top panel) and expression of CD33 and CD14 antigens (bottom panel) of cells from 476081 after drug treatment. (G) Heatmap showing mRNA fold change using the nanostring platform of drug treated cells relative to mock. Up-regulated genes are shown in red; down-regulated genes in green. Experiments were performed in triplicate; all data were normalized to GAPDH expression levels.

Characterization of UPN 476081. Cells from 476081 were grown on stroma for 4 days, followed by a 3-day treatment with cytarabine (AraC) or decitabine (DAC). Drug was administered daily. After 3 days of treatment, the number of viable cells (A), annexin-positive cells (B), and total 5-methycytosine content by LC-MS/MS (C) was determined. (D-E) cell cycle profiles were determined after overnight EdU incorporation and DNA content was measured by FxCycle Violet DNA dye. Shown are representative flow plots (D) and distributions of cell cycle phase after drug treatment (E). All experiments were performed in triplicate; error bars represent 95% confidence intervals; asterisk indicates a significant change from mock treatment. (F) CD45dim/SSlow blast gate (top panel) and expression of CD33 and CD14 antigens (bottom panel) of cells from 476081 after drug treatment. (G) Heatmap showing mRNA fold change using the nanostring platform of drug treated cells relative to mock. Up-regulated genes are shown in red; down-regulated genes in green. Experiments were performed in triplicate; all data were normalized to GAPDH expression levels.

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