Research Paper Volume 7, Issue 8 pp 553—567

Knockdown of Indy/CeNac2 extends Caenorhabditis elegans life span by inducing AMPK/aak-2

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Figure 1. (A) Representative survival curves for N2 (WT), fed with E. coli GC363 harboring the empty vector L4400 (empty vector control) versus E. coli GC363 harboring INDY/CeNAC2-specific siRNA. INDY/CeNAC2-leads to a significant life span extension (P < 0.001). Life span experiments were repeated four times in independent experiments, figure shows the second experiment, other data in Table 1.(B) Oil Red O staining of N2 wild type fed with E.coli GC363 harboring the empty vector L4400 (empty vector control) versus E.coli GC363 harboring INDY/CeNAC2-specific siRNA (C). N2 wild type worms showed more intensive staining and higher total area stained compared to RNAi treatment (D), Densitometric analysis of B and C, **p = 0.005. (E) Immunoblot analysis of different C. elegans strains (N2, TG) and fasted mouse liver lysate (C) as positive control for phosphorylation (activation) of AMPK/ aak-2 after knockdown of INDY/CeNAC2 (N2/I). Down regulation of INDY/CeNAC2 leads to activation of AMPK / aak-2 compared to control fed wildtype N2. Negative control TG38 (TG) with missing phosphorylation site for AMPK-Thr172 shows no activation when fed with empty vector control neither when treated with RNAi against INDY/CeNAC2 (G/i). Control (c) = liver lysates from fasted mice.