Figure 7
Figure 7. Association between transcription and translation inhibition and cell death. (A) Global RNA synthesis inhibition versus apoptosis levels and its linear regression. RNA synthesis was measured in percentages and apoptosis was defined by annexin V positivity. Values were taken from dose-dependent SGI-1776 treatments in JeKo-1, Mino, SP-53, and Granta-519 cell lines. In experiments performed in JeKo-1 and Mino after dose-dependent SGI-1776 treatments for 24 hours, the following plots were generated: c-Myc (Ser62) phosphorylation versus global RNA synthesis, where c-Myc (Ser62) levels were normalized to total c-Myc to obtain the ratios (B); c-Myc (Ser62) phosphorylation versus MCL1 mRNA expressions, where c-Myc (Ser62) levels were normalized to total c-Myc and MCL1 mRNA was normalized to 18S rRNA controls to obtain the ratios (C); inhibition of 4E-BP1 (Thr37/46) phosphorylation versus apoptosis, where 4E-BP1 (Thr37/46) phosphorylation levels were normalized to total 4E-BP1 to obtain the ratios (D); and 4E-BP1 (Thr37/46) phosphorylation versus Mcl-1 protein expression, where phospho–4E-BP1 (Thr37/46) was normalized to total 4E-BP1 protein and Mcl-1 protein was normalized to GAPDH levels (E). Linear regression was performed, and r2 values and P values were calculated using the GraphPad Prism Version 5 software.

Association between transcription and translation inhibition and cell death. (A) Global RNA synthesis inhibition versus apoptosis levels and its linear regression. RNA synthesis was measured in percentages and apoptosis was defined by annexin V positivity. Values were taken from dose-dependent SGI-1776 treatments in JeKo-1, Mino, SP-53, and Granta-519 cell lines. In experiments performed in JeKo-1 and Mino after dose-dependent SGI-1776 treatments for 24 hours, the following plots were generated: c-Myc (Ser62) phosphorylation versus global RNA synthesis, where c-Myc (Ser62) levels were normalized to total c-Myc to obtain the ratios (B); c-Myc (Ser62) phosphorylation versus MCL1 mRNA expressions, where c-Myc (Ser62) levels were normalized to total c-Myc and MCL1 mRNA was normalized to 18S rRNA controls to obtain the ratios (C); inhibition of 4E-BP1 (Thr37/46) phosphorylation versus apoptosis, where 4E-BP1 (Thr37/46) phosphorylation levels were normalized to total 4E-BP1 to obtain the ratios (D); and 4E-BP1 (Thr37/46) phosphorylation versus Mcl-1 protein expression, where phospho–4E-BP1 (Thr37/46) was normalized to total 4E-BP1 protein and Mcl-1 protein was normalized to GAPDH levels (E). Linear regression was performed, and r2 values and P values were calculated using the GraphPad Prism Version 5 software.

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