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. Author manuscript; available in PMC: 2021 Mar 8.
Published in final edited form as: Science. 2020 Dec 10;371(6528):eabc8059. doi: 10.1126/science.abc8059

Figure 1. Elevated activity of PGE2 degrading enzyme, 15-PGDH, is associated with aging, and 15-PGDH knockdown in aged muscle tissues increases strength.

Figure 1.

(A) PGE2 and PGD2 catabolism scheme. (B) Prostaglandin levels in Tibialis anterior (TA) muscles quantified by LC-MS/MS (n=14 young, n=8 aged). (C) Representative chromatogram of the PGE2 and PGD2 levels analyzed by LC-MS/MS in young (left) and aged (right) TA muscles. (D) 15-PGDH specific enzymatic activity assayed in tissues of young and aged mice. Activity is expressed as percent change relative to young. (n=4–5 young, n=3–5 aged). (E) 15-PGDH (Hpgd) RNAseq expression data from young (n=4) and aged (n=5) mouse muscles. TPM, transcripts per million. (F) 15-PGDH immunoblots from young and aged muscle lysates (n=4 each). (G-Q) Intramuscular (i.m.) injection of AAV9 carrying a construct of an shRNA against 15-PGDH (sh15PGDH) or scramble (scr) control into the TA and Gastrocnemius (GA) of young and aged mice. (G) Experimental scheme. (H) Expression levels of 15-PGDH in scr and sh15PGDH infected muscles and young control (n=5 young, n=4 aged scr, n=5 aged sh15PGDH). (I) 15-PGDH specific enzymatic activity assayed in muscle tissues of sh15PGDH infected aged muscles normalized to scr (n=6 aged scr, n=5 aged sh15PGDH). (J) Prostaglandin levels in GA muscles quantified by LC-MS/MS (n=5 young scr, n=4 young sh15PGDH, n=3 aged scr, n=4 aged sh15PGDH). (K) Representative GA cross-section of scr and sh15PGDH infected aged muscles. DAPI, blue; LAMININ, green. Bar=50 µm. (L) Myofiber cross-sectional areas (CSA) in scr and sh15PGDH infected aged GAs (n=7 aged scr, n=8 aged sh15PGDH). (M) Mean CSA (n=4 young scr, n=3 young sh15PGDH, n=7 aged scr, n=8 aged sh15PGDH). (N) Mass of dissected TA. (O) Mass of dissected GA. (P) Plantar flexion tetanic force (absolute values). (Q) Plantar flexion tetanic force (relative to baseline). (N-Q): n=8–10 young scr, n=8–9 young sh15PGDH, n=12–14 aged scr, n=14 aged sh15PGDH. *P<0.05, **P<0.01, ****P<0.0001. ANOVA test with Bonferroni correction (N-Q) or Fisher’s LSD (B,H,J,L,M) for multiple comparisons; unpaired t-tests (D-F,I). Means±s.e.m. Abbreviations: PGEM, 13,14-dihydro-15-keto-PGE2; Spl. Spleen; Mus. Muscle; mo. months; i.m. intramuscular. C57Bl/6 young (2–4 mo.) and aged (>24 mo.) mice were used.