Fig. 4

Effect of the addition of pharmacological inhibitors of PI3K/PTEN/Akt and mTOR signalling pathways during cryopreservation and/or in vitro organotypic culture on follicle activation of neonatal ovaries. A and B Ratio of phosphorylated to total protein Akt and rps6. C Representative images of p-Akt (a-c) and p-rps6 (d-f) staining for the different groups. SF = slow-frozen/thawed ovaries with or without inhibitors followed or not by organotypic culture (cult) of 24 h with or without inhibitors. CT = control; LY = LY294002; Ra = rapamycin. Scale bar represents 50 µm. 3–5 experimental replicates were performed. Ovaries from C57Bl/6 mice, 3–7 days old; n = 24–40 ovaries per group