Fig. 4: R199C is a stronger follicle development inducer than AARS2.

A–F AARS2 inhibits the TCA cycle and accumulates anabolites in GCs. Heatmap of annotated metabolites in TCA cycle (A), glycolysis (B), and PPP (C). The inhibition of the TCA cycle is further confirmed by measuring OCR (D), ATP (E), and lactate production (F) in GCs of C57 and GOE mice. G–L R199C has a strong ability to reprogram GC metabolism. The metabolites changes in TCA cycle (G), glycolysis (H), PPP (I), and OCR (J), ATP (K), and lactate (L) were compared between AARS2^−/− COV434 cells and AARS2^−/− COV434 cells expressing either AARS2 or R199C. M AARS2 activates mTORC1 signaling in COV434 cells. p-S6K and p-4EBP were detected in COV434 cells, AARS2^−/− COV434 cells, AARS2^−/− COV434 cells expressing AARS2, and AARS2 mutants. N AARS2 promotes cell proliferation in COV434 cells. CCK-8 assay was employed to measure growth of COV434, AARS2^−/− COV434 cells, and AARS2^−/− COV434 expressing AARS2 or R199C. O AARS2 activates mTORC1 signaling in mouse GCs. p-S6K and p-4EBP were detected in C57 and AARS2 GOE mouse GCs. P AARS2 promotes cell proliferation in mouse GCs. CCK-8 assay was employed to measure growth of GCs from C57 and AARS2 GOE mice. Q AARS2 promotes mouse follicular development. Representative images of PD8-C57 and AARS2 GOE mice ovarian sections, stained for DAPI, FOXL2 (right panel). The secondary follicles in 6 mice ovaries were counted (left panel) (scale bars, 200 μm (top panel), 20 μm (bottom panel)).