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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

We incubated cells with TG101209 (5μM) and monitored for differences in signaling and apoptotic proteins. A) MM1S cells and B) RPMI 8226 cells were treated with TG101209 (5μM) for 1, 2, 4 or 8 hours. We observed down regulation of pJak2 and pStat3 with no change in total Jak2 and Stat3 levels. β-actin was used as control. C) Patient 1 and D) Patient 2 samples were treated with TG101209 (5μM) for 4 or 8 hours. We observed down regulation of pJak2 and pStat3 with no change in total Jak2 and Stat3 levels. β-actin was used as control. E) MM1S cells and F) RPMI 8226 cells were treated with TG101209 (5μM) for 1, 2, 4 or 8 hours and performed western blotting for Bcl2, Bcl-xl, Mcl1, Xiap, pAkt and pErk. We observed down regulation of Bcl-xl in both cell lines. Mcl1 was down regulated in RPMI 8226 cells and Xiap levels were decreased in both cell lines post drug treatment. We observed up-regulation of pAkt and pErk protein levels post treatment with the drug indicating cross talk between pathways. β-actin was used as control. G) Patient 1 and H) Patient 2 samples were treated with TG101209 (5μM) for 4 or 8 hours and performed western blotting for Bcl2, Bcl-xl, Mcl1, Xiap, pAkt and pErk. We observed down regulation of Xiap in both patient samples post drug treatment. Mcl 1 was down regulated in patient 1. pAkt was down regulated at 4 hrs which was abolished at 8 hrs of drug treatment. pErk levels were up regulated following drug treatment. Bcl2 and Bcl-xl levels did not show any observable changes. Bcl2, BclXl and Mcl1, pAkt showed down regulation post drug treatment in patient 2. There was no significant difference in pErk levels in patient 2.

We incubated cells with TG101209 (5μM) and monitored for differences in signaling and apoptotic proteins. A) MM1S cells and B) RPMI 8226 cells were treated with TG101209 (5μM) for 1, 2, 4 or 8 hours. We observed down regulation of pJak2 and pStat3 with no change in total Jak2 and Stat3 levels. β-actin was used as control. C) Patient 1 and D) Patient 2 samples were treated with TG101209 (5μM) for 4 or 8 hours. We observed down regulation of pJak2 and pStat3 with no change in total Jak2 and Stat3 levels. β-actin was used as control. E) MM1S cells and F) RPMI 8226 cells were treated with TG101209 (5μM) for 1, 2, 4 or 8 hours and performed western blotting for Bcl2, Bcl-xl, Mcl1, Xiap, pAkt and pErk. We observed down regulation of Bcl-xl in both cell lines. Mcl1 was down regulated in RPMI 8226 cells and Xiap levels were decreased in both cell lines post drug treatment. We observed up-regulation of pAkt and pErk protein levels post treatment with the drug indicating cross talk between pathways. β-actin was used as control. G) Patient 1 and H) Patient 2 samples were treated with TG101209 (5μM) for 4 or 8 hours and performed western blotting for Bcl2, Bcl-xl, Mcl1, Xiap, pAkt and pErk. We observed down regulation of Xiap in both patient samples post drug treatment. Mcl 1 was down regulated in patient 1. pAkt was down regulated at 4 hrs which was abolished at 8 hrs of drug treatment. pErk levels were up regulated following drug treatment. Bcl2 and Bcl-xl levels did not show any observable changes. Bcl2, BclXl and Mcl1, pAkt showed down regulation post drug treatment in patient 2. There was no significant difference in pErk levels in patient 2.