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. Author manuscript; available in PMC: 2015 Sep 25.
Published in final edited form as: Cell Rep. 2014 Sep 4;8(6):1767–1780. doi: 10.1016/j.celrep.2014.08.006

Figure 5. Intestinal AMPK activation maintains intestinal homeostasis during aging and extends lifespan.

Figure 5

(A) Survival curves of TIGS-2>UAS-mCh-AMPK females with or without RU486-mediated transgene induction (p<0.0001; log-rank test; n>116 flies).

(B) Intestinal integrity during aging in TIGS-2>UAS-mCh-AMPK females with or without RU486-mediated transgene induction (p<0.01, at 30 days, p<0.05 at 45 days; binomial test; n>127 flies/condition).

(C) Expression levels of autophagy genes in from intestines of TIGS-2>UAS-mCh-AMPK female flies at 10 days of adulthood with or without RU486-mediated transgene induction. (t-test; n>3 of RNA extracted from 15 intestines/replicate).

(D) GFP-Atg8a staining. Representative images of enterocytes from the posterior midgut of 10 day old TIGS-2>UAS-mCh-AMPK, pGFP-Atg8a females with or without RU486-mediated transgene expression (red channel-TO-PRO-3 DNA stain, green channel-GFP-Atg8a, scale bar represents 10µm).

(E) Quantification of posterior midgut GFP-Atg8a foci (p<0.0001; t-test; n>10 confocal stacks from posterior midgut/condition; one fly per replicate stack).

(F) Lysotracker Red staining. Representative images of posterior midgut enterocytes from 10 day old TIGS-2>UAS-AMPK females with or without RU486-mediated transgene induction (scale bar represents 10µm).

(G) Quantification of acidophilic vesicles (p<0.0001; t-test; n>10 confocal stacks from posterior midgut/condition; one fly per replicate stack).

(H) Survival curves without food of TIGS-2>UAS-mCh-AMPK females with or without RU486-mediated transgene induction (p<0.001; log-rank; n>257 flies).

(I) Body mass during starvation of TIGS-2>UAS-mCh-AMPK females with or without RU486-mediated transgene induction (p<0.05, at 48hours and p<0.01 at 96 hours of starvation; t-test; n>6 samples/condition; 10 flies weighed/sample).

(J) Whole body lipid stores during starvation of TIGS-2>UAS-mCh-AMPK females with or without RU486-mediated transgene induction (p<0.01 at 48 hours, and p<0.001 at 96 hours of starvation; t-test; n>3 samples/condition/timepoint; lipids extracted from 5 flies/sample).

Data are represented as mean ± SEM. RU486 was provided in the media after eclosion at a concentration of 25µg/ml (E, F) and 100µg/ml for all other figures.