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. 2007 Jul 2;178(1):15–22. doi: 10.1083/jcb.200612017

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Copyright © 2007, The Rockefeller University Press

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Figure 4. — CERT regulates PKD activation and secretory transport. (A) HEK293T cells transiently expressing CERT-WT and -S132A were lysed, and PKD activation was analyzed by immunoblotting with pS910 PKD antibody (top). Equal loading was verified by reprobing with PKD1-specific antibody (middle). The expression of CERT proteins was verified by immunoblotting with GFP-specific antibody (bottom). (B and D) HEK293T cells were transfected with the indicated expression plasmids (B), and COS7 cells were transfected with the indicated siRNAs (D) together with ssHRP-Flag plasmid as described in Materials and methods. The medium was analyzed for HRP activity after 0, 1, 3, and 6 h by chemiluminescence. Values correspond to the mean of triplicate samples, and error bars represent SEM. RLU, relative light units. (C) COS7 cells were transfected with the siRNAs indicated, and CERT expression was analyzed after 72 h by immunoprecipitation and Western blotting using a CERT-specific antibody (top). Tubulin levels were not affected (bottom). CERT is marked with an arrow.