Fig. 3.

PIM1 interacts with NFATC1 in PC-3 cells. a Wild-type (WT) or multi mutant (MM) NFATC1 were transiently expressed in PC-3 cells and their subcellular localization patterns were analysed by confocal microscopy after staining with anti-Flag antibody. Shown are means from one experiment with three parallel samples. b The physical interactions between RFP-tagged PIM1 and GFP-tagged WT or MM NFATC1 proteins were analysed by fluorescence-lifetime imaging microscopy (FLIM) from samples of transiently transfected PC-3 cells. 24 h after transfection, cells were treated overnight with DMSO or 10 μM DHPCC-9. Shown on the left are representative images of negative control cells with expression of empty GFP or RFP vectors, while on the right are single channel or merged images of cells co-transfected with GFP- or RFP-tagged vectors. Scale bar 20 μm. c Shown are average GFP lifetimes from two independent FLIM experiments along with sample numbers inside the black bars as well as representative images with a heatmap. Lowest negative control (GFP + RFP) value was set as the limit for physical interaction