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. Author manuscript; available in PMC: 2020 Feb 19.
Published in final edited form as: Nat Metab. 2019 Aug 19;1(8):790–810. doi: 10.1038/s42255-019-0097-9

Figure 4. Adult GSC proliferation determines cold-induced longevity.

Figure 4

a, BrdU staining of germ lines from wild-type worms treated with 100 μg ml-1 FUdR during adulthood at the indicated temperatures after development at 20°C. Worms were examined at day 5 of adulthood. Nuclei were stained with DAPI. Scale bar represents 20 µm. The images are representative of 3 independent experiments. b, FUdR during adulthood does not affect lifespan at 20°C (20°C mean ± s.e.m: 18.23 ± 0.49, 20°C + FUdR: 18.28 ± 0.40, P= 0.8617), but reduces cold-induced longevity (15°C: 29.08 ± 0.60, 15°C + FUdR: 17.49 ± 0.40, P<0.0001). c, BrdU staining of germ lines from wild-type worms examined at day 5 of adulthood. Scale bar represents 20 µm. The images are representative of 3 independent experiments. d, Percentage of BrdU-positive cells/total nuclei (mean ± s.e.m., Vector/mog-5/iff-1 RNAi (n= 33 germ lines), pgl-1/iff-2/eif-2A RNAi (n= 36), germ lines from 3 independent experiments). Analysis by two-tailed Student’s t-test for unpaired samples (**** (P<0.0001)). e, Knockdown of mog genes shortens cold-induced longevity (15°C) in wild-type worms. Vector RNAi mean ± s.e.m: 26.60 ± 0.74, mog-5: 20.52 ± 0.63 (P<0.0001); mog-4: 21.83 ± 0.69 (P<0.0001); mog-1: 22.87 ± 0.76 (P= 0.0020). f, mog RNAi does not reduce lifespan of wild-type worms at 20°C. Vector RNAi mean ± s.e.m: 17.96 ± 0.46, mog-5: 17.13 ± 0.28 (P= 0.0653), mog-4: 17.68 ± 0.34 (P= 0.3608), mog-1: 17.58 ± 0.34 (P= 0.2567). g, mog RNAi does not shorten lifespan of wild-type worms at 25°C. Vector RNAi mean ± s.e.m: 13.50 ± 0.42, mog-5: 13.93 ± 0.27 (P= 0.7344), mog-4: 14.88 ± 0.27 (P= 0.0420), mog-1: 14.76 ± 0.36 (P= 0.0685). h, Knockdown of pgl and glh genes does not suppress cold-induced longevity in wild-type worms. Vector RNAi mean ± s.e.m: 30.35 ± 0.85, mog-5: 24.92 ± 0.44 (P<0.0001), pgl-1: 30.96 ± 0.65 (P= 0.6424), pgl-3: 28.95 ± 0.72 (P= 0.2590), glh-1: 29.64 ± 0.94 (P= 0.9853), glh-4: 29.57 ± 0.81 (P= 0.7474). i, Loss of iff-1 shortens cold-induced longevity in wild-type worms at 10°C. Vector RNAi mean ± s.e.m: 42.45 ± 0.84, iff-1: 35.52 ± 0.54 (P<0.0001), iff-2: 43.00 ± 0.66 (P= 0.6186), eif-2A: 43.11 ± 0.68 (P= 0.8526). j, iff-1 RNAi reduces cold-induced longevity in wild-type worms at 15°C. Vector RNAi mean ± s.e.m: 29.19 ± 0.86, iff-1: 21.66 ± 0.76 (P<0.0001), iff-2: 29.24 ± 0.85 (P= 0.8227), eif-2A: 28.53 ± 0.74 (P= 0.3853). k, iff-1 RNAi does not reduce lifespan at 20°C. Vector RNAi: mean ± s.e.m: 19.33 ± 0.45, iff-1: 19.79 ± 0.51 (P= 0.1317), iff-2: 19.61 ± 0.50 (P= 0.2137), eif-2A: 18.65 ± 0.44 (P= 0.2061). l, iff-1 RNAi induces a moderate lifespan extension at 25°C. Vector RNAi mean ± s.e.m: 13.23 ± 0.40, iff-1: 15.36 ± 0.39 (P<0.0001), iff-2: 13.69 ± 0.42 (P= 0.4603), eif-2A: 13.19 ± 0.33 (P= 0.6935). In lifespan experiments, P-values: two-sided log-rank test, n= 96 worms/condition. RNAi initiated at day 1 of adulthood in all the experiments. Supplementary Data 3 contains replicate data of lifespan experiments.