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Fig. 3. SCA14 mutations located in C1B enhanced PKC ${gamma}$ translocation kinetics upon phorbol ester stimulation. (A) Confocal images of HeLa cells cotransfected with wild-type PKC ${gamma}$ -GFP and SCA14 mutant (G118D, V138E and C142S) PKC ${gamma}$ -RFP. PMA activation induces irreversible translocation of PKC ${gamma}$ to the plasma membrane. Images shown were recorded 140 and 440 seconds after addition of 400 nM PMA. Scale bars: 10 µm. (B) Translocation analysis showed that PMA (400 nM) stimulation resulted in an increase in translocation kinetics of all three SCA14 mutant PKC ${gamma}$ proteins compared with wild-type PKC ${gamma}$ to the plasma membrane. (C) Translocation analysis of wild-type PKC ${gamma}$ -GFP, SCA14 mutant PKC ${gamma}$ -GFP (G118D, V138E and C142S) and ${Delta}$ VPKC ${gamma}$ -GFP upon PMA (400 nM) stimulation. Each of the traces in B and C represent mean ± s.e.m. of 4-6 cells from at least three independent experiments.

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