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. 2023 Sep 16;26(10):107960. doi: 10.1016/j.isci.2023.107960

Figure 1.

Figure 1

SQST-1 dynamics and lifespan modulation is temperature-dependent in C. elegans

(A–F) Lifespan analysis of wild-type animals and animals over-expressing SQST-1 fused to RFP, GFP or GFP::RFP fed OP50 E. coli, developmentally raised at 20°C and then grown at 25°C (A–C) or 20°C (D–F) during adulthood (n = 100). Insets include corresponding representative images of transgenic animals at Day 5 of adulthood, and quantified fluorescence, ±SD t-test ∗∗∗∗p < 0.001.

(G and H) (G) sqst-1 mRNA levels in wild-type and transgenic animals were quantified by qPCR. Biological triplicates ±SD t-test ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (H). Levels of GFP and RFP were measured in transgenic tandem SQST-1:GFP::RFP animals (leveraging the pH sensitivity of GFP to visualize SQST-1 accumulation in autolysosome) after incubating Day 1 animals at 20°C, 25°C or 30°C for 24 h. Average of 10 worms per image. n = 5 images per condition ±SD ANOVA ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Difference in signal between GFP and RFP is due to the low pH sensitivity of GFP. Thus, RFP-only signal represents SQST-1 protein in autolysosomes.

(I) Representative images of transgenic animals (quantified in h.) incubated at 20°C, 25°C or 30°C for 24 h. Details about lifespan analyses and repeats are available in Table S4, Mantel-Cox log rank. n.s.: not significant, ∗∗p < 0.01,∗∗∗∗p < 0.001.