Figure 6. Notch1 upregulates Pim protein levels.

A. The stably transfected MCF-7/N1ΔE cells were treated with increasing amounts of doxycycline to induce Notch1 overexpression, after which N1ICD and Pim protein levels were measured by Western blotting. B. PC-3 cells were transiently transfected with wild-type (WT), phosphodeficient (SA) or phosphomimicking (SE) N1ΔE, and 48 hours later, the N1ICD and Pim protein levels were analysed by Western blotting. C. Untransfected PC-3 cells were treated with 5 μg/ml of DAPT for 24 h prior to Western blotting with Pim antibodies. D. Untransfected PC-3 cells were treated with 10 μM DHPCC-9 for 24 h prior to Western blotting with antibodies that specifically recognise cleaved N1ICD. Similar experiments were repeated at least twice.