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. Author manuscript; available in PMC: 2009 Oct 1.
Published in final edited form as: Cancer Res. 2008 Oct 1;68(19):8156–8163. doi: 10.1158/0008-5472.CAN-08-1061

Figure 1. GSK-3 is required for constitutive NF-κB DNA binding.

Figure 1

EMSA was performed on Panc-1 and MiaPaCa-2 nuclear extracts using 32P-labeled NF-κB-specific probe. (A) Super-shift analysis was performed using antibodies against p65 and p50. Arrows indicate p65/50 and p50/p50 NF-κB complexes. Non-specific binding is specified by NS. (B) Cells were treated with vehicle control (DMSO) or GSK-3 inhibitors (AR-A014418 and SB216763) at 15 and 25μM for 24 hours. (C) Panc-1 cells were transiently transfected with 100nM siRNA targeted against GSK-3α, GSK-3β, combined GSK-3α/β, and non-targeting control (siCTRL) for 48 hours. Cytoplasmic extracts were harvested and separated by SDS-PAGE. Immunoblots were performed using the specified antibodies. (D) Nuclear extracts were harvested from Panc-1 and MiaPaCa-2 cells transfected with siRNA as described above and EMSA was performed.