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Fig. 3. (A) TPA-induced downregulation of PKC{alpha} is sensitive to a proteasome inhibitor. Control (empty-vector-transfected) RBL cells and RBL-Syt II- cells were treated for 12 hours with TPA (50 nM) in the absence or presence of the lysosomal inhibitor (leupeptin, 10 µg/ml) or the proteasomal inhibitor (ALLN, 38.5 µg/ml). Cytosolic and membranal fractions were separated as described in the Materials and Methods, and the amount of PKC{alpha} in each fraction was determined as described in Fig. 1. Data represent one of two separate experiments. (B) TPA-induced downregulation of PKC{alpha} is sensitive to the PKC inhibitor Go 6976. Control (empty vector transfected) RBL cells and RBL-Syt II- cells were treated for 12 hours with TPA (50 nM) in the absence or presence of the PKC inhibitor Go 6976 (100 nM). The cellular level of PKC{alpha} was determined as described in Fig. 1. Data represent one of three separate experiments.