Profiling of Luteal Transcriptome during Prostaglandin F2-Alpha Treatment in Buffalo Cows: Analysis of Signaling Pathways Associated with Luteolysis
Figure 4
Effects of PGF2α on expression of LH/CGR and downstream signaling molecules, Akt and FKHR.
(A) Protein levels of LH/CGR in bovine CL. Protein lysate (100 µg) prepared from CL tissue collected before (0 h) and post (3, 6 and 18 h) PGF2α treatment were resolved on 10% SDS PAGE, transferred onto PVDF membrane and immunoblot analysis was performed using anti-LHCGR and anti-β-actin antibody. A representative immunoblot for each of the antibody probed is shown. The immunoblot probed with β-actin antibody indicates loading control for each lane. Densitometric values were determined and indicated as mean±SEM (n = 3 animals/time point), relative to intensity of β-actin for each time point post PGF2α treatment. The values of immunoblot analysis has been put on top of each lane and lane with different alphabets indicate statistical significance, p<0.05. (B) PKA activity in the bovine CL before (0 h) and after (3 and 18 h) PGF2α administration. Values represent mean±SEM for each time point post PGF2α administration (n = 3 animals/time point). Individual bars with different alphabets indicate statistical significance (p<0.05). (C, D and E) Protein levels of pCREB, CREB, pAkt, Akt, pFKHR and FKHR in the CL. Protein lysate (50 µg) of each CL tissue collected at different time points was subjected to immunoblot analysis employing anti-pCREB, anti-CREB, anti-pAkt, anti-Akt, anti-pFKHR, anti-FKHR antibodies. A representative immunoblot for each of the antibody probed is shown. Densitometric values were determined and represented as mean±SEM (n = 3 animals/time point), relative to intensity of total CREB (C), Akt (D) and FKHR (E) for each time point post PGF2α treatment. The values of immunoblot analysis has been put on top of each lane and lanes with different alphabets indicate statistical significance (p<0.05).