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Fig. 8. Differential activation (phosphorylation) pattern of AMPK in differentiating wild-type (ple +/+) and plectin-deficient (ple -/-) myocytes: specificity and phosphatase independence. (A,B) Total cell lysates of wild-type or plectin-deficient myocytes, differentiated (diff.) for the time periods indicated (0-6 days), were subjected to immunoblotting using antibodies to unphosphorylated/phosphorylated (total) and phosphorylated (P-) forms of AMPK and p38 MAP kinase, respectively. (C,D) Signal intensities of phosphorylated AMPK and p38 MAP kinase protein bands were densitometrically determined in three independent experiments (including those shown in A,B), and normalised to total AMPK and p38 MAP kinase, respectively. Error bars represent the s.e.m. (E-G) Enzymatic activities of phosphatases (PP) 1, 2A, and 2C were measured in cell lysates obtained from undifferentiated myoblasts (E), or myoblasts differentiated for 3 (F) or 6 days (G), using a fluorescent substrate. Error bars represent the s.e.m. of three independent experiments. (C-G) Differences between values in ple (+/+) and ple (-/-) were determined using an unpaired Student's t-test; ^*P<0.05.

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