Figure 4. Notch1 colocalizes and interacts with Pim1.

A. PC-3 cells were transiently transfected with GFP-tagged wild type (WT), phosphodeficient (SA) or phosphomimicking (SE) N1ICD, RFP-tagged Pim1 or empty GFP or RFP vectors. 24 h after transfection, cells were treated overnight with 0.1% DMSO, 10 μM DHPCC-9 or 5 μg/ml of DAPT. Fixed samples were analysed by confocal microscopy. Shown are representative single channel or merged images of DMSO-treated samples. B. Physical interactions between Pim1 and N1ICD were measured by fluorescence-lifetime imaging microscopy (FLIM). Samples expressing only GFP or GFP-tagged N1ICD were used as negative controls. Shown are average GFP lifetimes from two independent experiments along with analysed cell numbers inside the bars. C. Representative images from FLIM analyses.