The metabolic shift of late prepubertal spermatogonia is associated with AKT-pathway activation, expression of PLZF and epigenetic changes. A, Late prepubertal spermatogonia show higher expression of ACSS-2 and PLZF, downregulation of C-Kit compared to early prepubertal spermatogonia (mean ± SEM log fold change compared to mean Δct of early prepubertal spermatogonia; statistical analysis performed on raw Δct, P < .05, n = 4; Table S1). B, Representative image of Western blot analysis of sorted early versus late prepubertal spermatogonia for GAPDH, GFRA1, RET, PLZF, Acetyl-H3 relative to HPRT1. C-G, Quantification early versus late prepubertal spermatogonia; mean ± SEM: (C) PLZF expression (P = .0027, n = 4). D, Phosphorylation of AKT (Ser 473) relative to pan AKT (p = .042, n = 4), E, pan H3 acetyl (P = .039, n = 4), F, RET (P = .125, n = 4) and G, GFRA1 (P = .324, n = 4) (two-tailed, unpaired Student’s t test)