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. Author manuscript; available in PMC: 2011 Sep 1.

Published in final edited form as: Am J Hematol. 2010 Sep;85(9):675–686. doi: 10.1002/ajh.21785

TG101209 treatment (5μM) leads to accumulation of cells in G2/M stage of cell cycle. We treated A) MM1S or B) RPMI 8226 cells with TG101209 (5μM) for either 6, 12 or 24 hours and observed time dependent increase of cells in G2/M stage indicating growth arrest. Time of incubation with TG101209 is indicated on the X-axis and percent cells are indicated on the Y-axis. Next, we examined changes in expression levels of cell cycle related proteins post treatment with TG101209. We treated C) MM1S or D) RPMI 8226 cells with TG101209 (5μM) for 1, 2, 4 or 8 hours. Post treatment we made lysates and checked for expression levels of Cdk2, Cdk4, p27 and p21. β-actin was used as a loading control. We show clear down regulation of Cdk2 and Cdk4 in both cell lines and increase in p21 and p27 in MM1S cells. We also incubated two patient derived CD138+ cells with 5μM of TG101209 for 4 or 8 hours. E) Patient 1 samples showed reduction in levels of Cdk4 with no observable change in p21. p27 was down regulated at 8 hrs of incubation with the drug. F) Patient 2 samples showed reduction in levels of Cdk2 and p27. p21 was up-regulated post drug treatment.

TG101209 treatment (5μM) leads to accumulation of cells in G2/M stage of cell cycle. We treated A) MM1S or B) RPMI 8226 cells with TG101209 (5μM) for either 6, 12 or 24 hours and observed time dependent increase of cells in G2/M stage indicating growth arrest. Time of incubation with TG101209 is indicated on the X-axis and percent cells are indicated on the Y-axis. Next, we examined changes in expression levels of cell cycle related proteins post treatment with TG101209. We treated C) MM1S or D) RPMI 8226 cells with TG101209 (5μM) for 1, 2, 4 or 8 hours. Post treatment we made lysates and checked for expression levels of Cdk2, Cdk4, p27 and p21. β-actin was used as a loading control. We show clear down regulation of Cdk2 and Cdk4 in both cell lines and increase in p21 and p27 in MM1S cells. We also incubated two patient derived CD138+ cells with 5μM of TG101209 for 4 or 8 hours. E) Patient 1 samples showed reduction in levels of Cdk4 with no observable change in p21. p27 was down regulated at 8 hrs of incubation with the drug. F) Patient 2 samples showed reduction in levels of Cdk2 and p27. p21 was up-regulated post drug treatment.