Figure 3
Figure 3. Effect of SGI-1776 in AML cell lines. (A) Inhibition of RNA synthesis in AML cell lines treated with SGI-1776. MV-4-11 (squares), MOLM-13 (triangles), and OCI-AML-3 (circles) lines were incubated with 0.1% DMSO alone, or 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours, then 1 hour before harvesting the cells, [3H]uridine was added to the cell culture as described in “RNA protein synthesis assay.” The results represent an average of triplicate experiments ± SEM. (B) Dose-dependent reduction of MCL-1 transcript levels in AML cell line treated with SGI-1776. MV-4-11 cells were treated with 0.1% vehicle DMSO alone, 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours, and the RNA was isolated. MCL-1 transcript levels were measured using real-time RT-PCR and normalized using 18S transcripts. Each RNA sample was assayed in triplicate and the results are expressed as a percentage of the MCL-1 expression level in cells treated with vehicle DMSO alone. The results represent an average of triplicate experiments ± SEM (C) Inhibition of protein synthesis in AML cell line treated with SGI-1776. MOLM-13 cells were incubated with 0.1% DMSO alone, or 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours, then 1 hour before harvesting the cells, [3H]leucine was added to the cell culture as described in “RNA protein synthesis assay.” The results represent an average of triplicate experiments ± SEM.

Effect of SGI-1776 in AML cell lines. (A) Inhibition of RNA synthesis in AML cell lines treated with SGI-1776. MV-4-11 (squares), MOLM-13 (triangles), and OCI-AML-3 (circles) lines were incubated with 0.1% DMSO alone, or 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours, then 1 hour before harvesting the cells, [3H]uridine was added to the cell culture as described in “RNA protein synthesis assay.” The results represent an average of triplicate experiments ± SEM. (B) Dose-dependent reduction of MCL-1 transcript levels in AML cell line treated with SGI-1776. MV-4-11 cells were treated with 0.1% vehicle DMSO alone, 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours, and the RNA was isolated. MCL-1 transcript levels were measured using real-time RT-PCR and normalized using 18S transcripts. Each RNA sample was assayed in triplicate and the results are expressed as a percentage of the MCL-1 expression level in cells treated with vehicle DMSO alone. The results represent an average of triplicate experiments ± SEM (C) Inhibition of protein synthesis in AML cell line treated with SGI-1776. MOLM-13 cells were incubated with 0.1% DMSO alone, or 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours, then 1 hour before harvesting the cells, [3H]leucine was added to the cell culture as described in “RNA protein synthesis assay.” The results represent an average of triplicate experiments ± SEM.

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