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. 2002 Jan 7;156(1):65–74. doi: 10.1083/jcb.200110047

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Copyright © 2002, The Rockefeller University Press

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Figure 4. — Localization of PKCμ-GFP at the Golgi compartment is required for phosphorylation of serines 738/742. (A) Differential phosphorylation of PKCμ-GFP deletion mutants. HEK293 cells were transfected with the indicated plasmids. Expression of the fusion proteins was monitored by Western blot analysis using an anti-GFP antibody. PKCμ-GFP phosphorylation was measured by phospho-specific antibodies recognizing phosphorylated Ser_738/742 and Ser₉₁₀. (B) Characterization of PKCμ-GFP phosphorylation mutants. (C) PKCμ_S738/742A-GFP colocalizes with the Golgi compartment–specific marker p24. (D) PKCμ-GFP with phosphorylated activation loop is exclusively recovered in the organelle fraction. HEK293 cells were transfected with PKCμ-GFP or PKCμ_K612W-GFP and separated into soluble proteins from organelles structures sedimenting at 100,000 g. Western blot analysis was performed by anti-GFP or phosphorylation-specific antibodies.

Figure 4. — Localization of PKCμ-GFP at the Golgi compartment is required for phosphorylation of serines 738/742. (A) Differential phosphorylation of PKCμ-GFP deletion mutants. HEK293 cells were transfected with the indicated plasmids. Expression of the fusion proteins was monitored by Western blot analysis using an anti-GFP antibody. PKCμ-GFP phosphorylation was measured by phospho-specific antibodies recognizing phosphorylated Ser_738/742 and Ser₉₁₀. (B) Characterization of PKCμ-GFP phosphorylation mutants. (C) PKCμ_S738/742A-GFP colocalizes with the Golgi compartment–specific marker p24. (D) PKCμ-GFP with phosphorylated activation loop is exclusively recovered in the organelle fraction. HEK293 cells were transfected with PKCμ-GFP or PKCμ_K612W-GFP and separated into soluble proteins from organelles structures sedimenting at 100,000 g. Western blot analysis was performed by anti-GFP or phosphorylation-specific antibodies.