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. 2023 Jan 18;14:288. doi: 10.1038/s41467-023-35952-z

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Lifespan showing that aak-2(ok524) abolishes GR-induced longevity (P = 1, P value determined by two-tailed Student’s t test). Results from one of three biological independent experiments are shown. b, c Lifespan showing that neuronal AAK-2 is required for GR-induced longevity. Tissue-specific RNAi animals are used for neuronal aak-2 RNAi knockdown (b) and intestinal aak-2 RNAi knockdown (c). Results from one of three biological independent experiments are shown. d Structure of aak-2⁺ gene and aak-2 isoform::gfp constructs. Coding regions are indicated as closed boxes; noncoding regions as line; introns spliced-out in DNA constructs as ∨. Locus of start codon for AAK-2 isoforms are indicated as dotted lines. e–g GFP image showing the expression of AAK-2 isoforms. GFP images are taken using aak-2(ok524);aak-2 isoform::gfp animals. AAK-2a is expressed in the head neurons (Ne) and excretory cells (Ex) (e). AAK-2c is expressed in multiple tissues, including the intestine (In), pharynx (Ph), muscles (Mu), hypodermis (Hy), and neuron (Ne) (f). AAK-2a/c transgenic animals show the combined expression patterns (g). Representative images obtained from at least four biological independent repeats. h Immunoblotting analysis showing the existence and relative levels of AAK-2 isoforms in the indicated animals. Worm lysates were detected with anti-GFP (top panel), and anti-α-tubulin (bottom panel). Representative images obtained from four biological independent repeats. i–k Lifespans showing contribution to GR-induced longevity by AAK-2 isoform(s). The lifespans of aak-2(ok524) expressing AAK-2 isoform(s) under endogenous promoter are shown; AAK-2a (i), AAK-2c (j) and AAK-2a/c (k). Results from one of three biological independent experiments are shown. l–n The lifespans of aak-2(ok524) transgenic animals expressing AAK-2a isoform in indicated tissues; AAK-2a in the neurons (l), the intestine (m) and the excretory cells (n). Results from one of three biological independent experiments are shown. Source data are provided as a Source Data file.