Figure S6.

Role of AMP Kinase and SKN-1 in Metformin-Induced Longevity, Related to Figure 6

(A) Metformin does not detectably alter pAAK-2 levels in 1-day adults when developing in the presence of different drug concentrations or in day-8 and day-16 adults.

(B) Metformin does not significantly increase lifespan in AMPK-deficient aak-2(ok524) mutant worms.

(C) Metformin decreases the exponential increase in age-related mortality in AMPK-deficient aak-2(ok524) mutant worms, but increases initial mortality.

(D) AMPK-deficient aak-2(ok524) mutant worms are hypersensitive to growth inhibition by phenformin as measured using the food clearance assay (Voisine et al., 2007).

(E) TMP does extend lifespan in aak-2(ok524) mutant worms, though the magnitude of the extension is slightly reduced.

(F) The folate-deficient (aroD) mutant E. coli (Virk et al., 2012) does extend lifespan in aak-2(ok524) mutants, though the magnitude of the extension is slightly reduced.

(G) sams-1 RNAi does not extend lifespan in aak-2(ok524) (AMPK-deficient) mutant worms.

(H) Metformin shortens lifespan in skn-1(zu135) mutant worms.

(I) Diagram showing AMPK-dependent expression of SKN-1 gene targets such as gst-4 by biguanides; after Onken and Driscoll (2010).

(J and K) Metformin induces gst-4::gfp expression in an AMPK dependent fashion, consistent with a previous report (Onken and Driscoll, 2010). (J) Representative epifluorescence microscopic images of gst-4::gfp expression in L4 larvae. (K) Quantification of reporter gene expression.

See Table S6 for statistics. Error bars, SEM of at least 3 independent biological replicates. ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001.