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. 2018 Jun 21;14(6):e1007379. doi: 10.1371/journal.pgen.1007379

Fig 3. Increased alg-1 is insufficient for lifespan extension.

Fig 3

(A) Survival curves of WT (black) and alg-2(ok304) (blue) strains transferred to vector control (solid) or alg-1 (dashed) RNAi at the L4 stage. ****P<0.0001 (log-rank). (B) Top: Western blot of ALG-1 protein levels in spe-9(hc88) and spe-9(hc88); alg-2(ok304) animals during L4 and adulthood. Bottom: Quantification of protein levels from four independent experiments. The samples were normalized to L4, and the error bars represent SEMs. **P<0.01, *P<0.05 (t-test). (C) Expression of endogenous ALG-1 tagged with GFP in WT versus alg-2(ok304) backgrounds visualized by fluorescence microscopy. Micrographs of heads & tails were captured at 40x magnification with 76 ms exposure. (D) Depiction of ALG-1 and miRNA binding sites in the alg-1 3’UTR detected by iCLIP and miRNA::target chimera analyses reported in Broughton et al., 2016 [32]. (E) Top: Western blot of ALG-1 protein levels expressed from the gene containing its native or a swapped 3’UTR during L4 and adulthood. Bottom: Quantification of protein levels from three independent experiments. The samples were normalized to WT L4, and the error bars represent SEMs. **P<0.01 (t-test). (F) Survival curves of the indicated strains, showing that replacement of the native alg-1 3’UTR with the Y45F10D.4 (swap) 3’UTR is not sufficient to produce a lifespan phenotype. Not significant (n.s.); ****P<0.0001 (log-rank).