MES-induced phenotypes are not observed in AMPK homolog <i>aak-2</i> mutant worms.

<p>(A–D) <i>aak-2</i> mutant worms (A, B: <i>gt33</i>; C, D: <i>ok524)</i> were treated once a day with MES (0.1 ms, 2 V/cm, 55 pps) for 20 min at larval stage. After the last treatment, worms were incubated under (A, C) heat stress (30°C) or (B, D) oxidative stress (4 mM paraquat) condition. Worms' survival was checked every 24 hr. Each time point is the average of three independent experiments using 80–100 animals per group. (E-H) <i>aak-2</i> mutant worms (E, F: <i>gt33</i>; G, H: <i>ok524</i>) worms were bred on NGM plates and treated once a day with MES for 20 min at larval stage. After the last treatment, total RNA was extracted and subjected to quantitative real-time PCR. The mRNA levels were normalized to the level of β-actin (internal control). Data are presented as mean ± SE. n.s. versus control, assessed by unpaired Student t-test. (I) <i>aak-2(gt33, ok524)</i> worms were bred on NGM plates and treated once a day with MES for 20 min at larval stage. After the last treatment, worms were stained with Nile red and observed using a fluorescent microscope. (J, K) Relative fluorescence intensity of Nile red for (J) <i>gt33</i> and (K) <i>ok524</i> was quantified. Data are representative of two independent experiments using 40–50 animals per group. Data are presented as mean ± SE. ^***P<0.001 versus control without D-glucose, n.s. versus control with D-glucose, assessed by one-way ANOVA. (L, M) <i>aak-2</i> mutant worms (L: <i>gt33</i>; M: <i>ok524</i>) were bred on NGM plates containing D-glucose (10 mM) and treated once a day with MES for 20 min at larval stage. After the last treatment, total RNA was extracted and subjected to quantitative real-time PCR. The mRNA levels were normalized to the level of β-actin (internal control). Data are presented as mean ± SE. n.s. versus control, assessed by unpaired t-test. n.s., not significant. For mRNA analysis (E–H and L–M), data shown are representative of two independent experiments using approximately 200 animals per group.