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. Author manuscript; available in PMC: 2011 May 1.
Published in final edited form as: Mol Cancer Ther. 2010 Apr 20;9(5):1158–1168. doi: 10.1158/1535-7163.MCT-09-1036

Figure 6.

Figure 6

Differential effects of BCL-2 and PIG-6 siRNAs on the apoptotic index and down stream proteins in OCI-AML3 cells. A and B. For transfection studies, OCI-AML3 cells were nucleofected with 10 nM non-targeting negative control, BCL-2 or PIG-6 siRNAs for 24 h. Cells were washed once with warm medium and then exposed to 10 nM 1,25D for 48 h followed by 10 µM nutlin-3a during the last 16 and 24 h. Cells were also exposed to single agents; 10 nM 1,25D for 48 h and 10 µM nutlin-3a for 16 and/or 24 h. The percentage of total fraction of Annexin V- positive cells was determined using the Guava Nexin™ Kit. The bars represent average values ± standard deviation of three independent experiments, with duplicate data points. P values were determined by Student t-test, where ** indicates p<0.05. C. For Western blotting, proteins extracted from nucleofected samples were resolved in 4–20% SDS-PAGE, transferred to PVDF membranes and incubated with the indicated antibodies. MEK1/2, ERK1/2 and β-actin proteins were the loading controls for the immunoblot.