Figure 3

LH ovoprotection against Cs is mimicked by cAMP increasing agents and requires PKA and AKT activities. (a) Percentage of healthy POs scored within the ovarian fragments after 24 h of culture: 10 μM Cs=30±3.3; 10 μM Cs+200 mIU LH=69±2.7; 10 μM Cs+1 mM 8Br-cAMP=47±2.1; 10 μM Cs+10 μM FRSK=70±2.5; 10 μM Cs+200 mIU LH+100 μM H89=46±1.8 or (b) 10 μM Cs=30±2.3; 10 μM Cs+200 mIU LH=74±4.4; 10 μM Cs+200 mIU LH 10 μM LY294002=30±2.7; 10 μM Cs+200 mIU LH+0.1 μM wortmannin=33±2.7. Statistical differences are versus Cs groups. (c) A representative western blotting (out of at least two performed in different samples) for AKT, p-AKT of P8 ovaries treated for 10, 20, 30 min with 200 mIU LH; incubation of the ovaries in 100 ng/ml KL is shown as a positive control. (d) A representative western blotting (out of at least two performed in different samples) for pERK1/2 and ERK1/2 of ovaries of the same age treated for 20 min with/out Cs and LH. (e) Densitometric analysis of pERK1/2 western blots. Statistical differences are versus not treated group. (f) Percentage of healthy POs scored within the ovarian fragments after 24 h of culture with/out Cs plus LH plus UO126: 10 μM Cs=34±3.5; 10 μM Cs+10 μM U0126=63±2.9; 10 μM Cs+200 mIU LH + 10 μM U0126=65±4.6. Statistical differences are versus Cs group