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. 2008 Jun 19;27(14):1919–1931. doi: 10.1038/emboj.2008.119

Figure 2.

Figure 2

Sin1 but not PDK1 is required for PKCα and Akt TM phosphorylation. (A) Phosphorylation of the TM and HM in PKCα and Akt, but not PKCδ and S6K, is eliminated in Sin1−/− MEF. Sin1+/+ or −/− MEF cells were cultured in normal medium (medium), serum-free medium for 12 h (starved) or restimulated with serum for 30 min (serum). Immunoblottings were performed using indicated antibodies. (B) Reintroduction of Sin1 in Sin1−/− MEF restores phosphorylation. Sin1−/− MEFs were infected with either empty vector or HA-Sin1-containing retroviruses. The cells were treated with or without 400 nM insulin for 30 min. (C) PDK1 is required for PKCα HM but not TM phosphorylation. Immunoblotting of lysates from PDK1+/+ and −/− ES cells was performed for PKCα and Akt phosphorylation. The cells were serum-starved for 3 h and then stimulated with or without 100 nM IGF for 30 min in the presence or absence of pretreatment with 100 nM wortmannin (W) for 30 min.