FIGURE 5.

Specific inhibition of PKCδ activity prevents its retention at mitochondria. A, COS-7 cells transfected with CKAR alone or together with wild-type PKCδ-RFP or with the gatekeeper mutant PKCδM425A-RFP were stimulated with 200 nm PDBu and then inhibited with 1 μm NaPP1 during FRET imaging. B and C, COS-7 cells co-transfected with δCKAR and either RFP, PKCδ-RFP, or PKCδM425A-RFP were either stimulated with 200 nm PDBu and then inhibited with 1 μm NaPP1 (B) or first inhibited with 1 μm NaPP1 and then stimulated with 200 nm PDBu (C). D, COS-7 cells co-transfected with Mito-CFP and either PKCδ-YFP or PKCδM425A-RFP were either left unpretreated/pretreated with Me₂SO vehicle control or pretreated with 1 μm NaPP1 for 30 min at 37 °C before 200 nm PDBu stimulation during FRET imaging. E, COS-7 cells co-transfected with CFP-targeted to the plasma membrane (PM-CFP) and either PKCδ-YFP or PKCδM425A-RFP were either left unpretreated/pretreated with Me₂SO vehicle control or pretreated with 1 μm NaPP1 for 30 min at 37 °C before 200 nm PDBu stimulation during FRET imaging. F, COS-7 cells co-transfected with δCKAR and PKCδ-RFP were pretreated with either Me₂SO vehicle control, 10 μm rottlerin, or 750 nm Gö6983 for 30 min at 37 °C before PDBu stimulation during FRET imaging.