Fig. 7: Downregulating lactate signal prevents POI traits in AARS2 GOE mice.

A β-alanine inhibits AARS2- and R199C-catalyzed PDHA1 and CPT2 lactylation in vitro. In vitro AARS2- and R199C-catalyzed PDHA1 (left) and CPT2 (right) lactylation were detected when β-alanine was present and absent in the reaction mix. B–C β-alanine inhibits AARS2- and R199C-induced mTORC1 signaling. p-S6K and p-4EBP (B) and cell proliferation (C, n = 3) were detected in COV434 cells and AARS2-overexpressing COV434 cells that were cultured with or without β-alanine. D–E AARS2 deletion abrogated β-alanine to inhibit GC mTORC1 and proliferation. The β-alanine effects on p-S6K and p-4EBP (D), and proliferation (E, n = 3) of COV434 cells and AARS2^−/− COV434 cells were detected. F–H β-alanine inhibits follicle development. The β-alanine effects on p-S6K and p-4EBP (F), GC proliferation (G, n = 3), and number of secondary follicles (H, n = 6) of C57 and AARS2 GOE mice were detected. See also Fig. S7. I–K β-alanine AARS2-dependently inhibits follicle development. The effects of β-alanine on p-S6K and p-4EBP (I), GC proliferation (J, n = 3), and number of secondary follicles of C57 and Aars2 GKO mice (K, n = 6) were detected. See also Fig. S7. L–N β-alanine relieves POI traits in AARS2 GOE mice. The effects of intraperitoneally injected β-alanine on estrous cyclicity (L) and the cycle phase times (M) of C57 and AARS2 GOE mice were detected, n = 6. (N) β-alanine increases AARS2 GOE mice litter size. Effects of intraperitoneally injected β-alanine on the cumulative number of pups per mother of C57 and AARS2 GOE mice were measured, n = 6. O β-alanine reserves primordial follicles and decreases developed follicles of AARS2 GOE mice. The follicle number of C57 and AARS2 GOE mice with or without intraperitoneal β-alanine injection was counted at PD3-, 5-, and 7-weeks, n = 6. See also Figs. S7 and S8.