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. 2007 Jun 27;405(Pt 2):331–340. doi: 10.1042/BJ20070299

Figure 4. PKC membrane redistribution by an ATP-competitive inhibitor requires its DAG-sensitive C1 domain.

Figure 4

(A) Whole-cell lysates were obtained from COS-7 cells transfected with the indicated plasmids using ice-cold homogenization buffer containing 1% Triton X-100 and subjected to SDS/PAGE, followed by immunoblotting with anti-GFP antibody. (B) HeLa cells transiently expressing GFP-tagged αΔC1A, δΔC1A and δΔC1B were treated with GF 109203X at 5 μM for 60 min (αΔC1A) or 1 μM for 20 min (δΔC1A and δΔC1B). A total of seven to nine cells were observed. (C) HeLa cells transiently expressing GFP-tagged αΔC1A, αΔC1B, δΔC1A and δΔC1B were treated with GF 109203X at 5 μM for 60 min (αΔC1A and αAC1B) or 1 μM for 20 min (δΔC1A and δΔC1B). A total of eight cells were observed. (D) HeLa cells transiently expressing GFP-tagged PKCα, αΔC1A, αΔC1B, PKCδ, δΔC1A and δΔC1B were treated with 25 or 50 μM DiC8 for 10 min. The translocation of δΔC1A indicates the decrease from the perinuclear region. (E) HeLa cells transiently expressing GFP-tagged ϵΔC1, ϵΔC1A, ϵΔC1B and ϵC259G were treated with 1 μM GF 109203X for 20 min. Results are representative of at least seven separate experiments.