Figure 1: NMD activity declines with age. | Nature Communications

Figure 1: NMD activity declines with age.

From: RNA surveillance via nonsense-mediated mRNA decay is crucial for longevity in daf-2/insulin/IGF-1 mutant C. elegans

Figure 1

(a) Diagram of a premature termination codon (PTC)-containing GFP-fused NMD reporter, sec-23p::gfp::lacZ(PTC) (lacZ(PTC)), and a control without PTC, sec-23p::gfp::lacZ(WT) (lacZ(WT)). Green boxes represent gfp exons, black boxes represent lacZ exons and grey boxes represent 3′ UTRs. (b) The images of young (Day 1) and old (Day 9) NMD reporter, lacZ(PTC) (scale bar, 100 μm). (c) Normalized fluorescence intensity of lacZ(PTC) to age-matched lacZ(WT) (n 23 from three independent experiments). The same bar graph was also used in Fig. 4i. (d) mRNA levels of the GFP-fused NMD reporter, lacZ(PTC), in young (Day 1) and old (Day 9) worms measured by qRT–PCR (n=2). (e) Normalized RT–PCR result of PTC-containing rpl-12 [rpl-12(PTC)] by rpl-12 [rpl-12(WT)], which do not contain PTC, transcripts (n=3). (f) lacZ(PTC) and lacZ(WT) were expressed from neuron-, hypodermis-, muscle- and intestinal-specific promoters. Normalized GFP intensities expressed from corresponding tissue-specific promoter-driven lacZ(PTC) to age-matched lacZ(WT) among different tissues during aging (n21 from three independent experiments). Neuron: rgef-1p::gfp::lacZ(PTC)/rgef-1p::gfp::lacZ(WT), hypodermis: dpy-7p::gfp::lacZ(PTC)/dpy-7p::gfp::lacZ(WT), muscle: myo-3p::gfp::lacZ(PTC)/myo-3p::gfp::lacZ(WT) and intestine: ges-1p::gfp::lacZ(PTC)/ges-1p::gfp::lacZ(WT). Hypodermal fluorescence intensity ratio of lacZ(PTC)/lacZ(WT) tended to be increased in Day 9 or Day 13 compared to that of Day 1, but the differences were not significant. Error bars represent s.e.m. (two-tailed Student’s t-test, *P<0.05, **P<0.01, ***P<0.001).

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