Fig. 4.

The S. pombe AMPK Ssp2 is activated by AMP. (A) Ssp2 immunoprecipitations followed by a kinase assay using SAMStide (a peptide containing the sequence around ACC S79 – an AMPK substrate) and pACC (S79) antibodies to detect Ssp2 activity. Following the enzyme reaction, the supernatant was used to perform a slot blot to detect pACC (S79) and the levels of Ssp2 kinase in reaction were determined by western blotting, 300 mM AMP was used. (B) Quantification of SAMStide phosphorylation under the conditions indicated. Results are mean±s.d., n=3. **P≤0.01 (two-tailed Student's t-test).