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Fig. 8 | Reproductive Biology and Endocrinology

Fig. 8

From: Docosahexaenoic acid (DHA) effects on proliferation and steroidogenesis of bovine granulosa cells

Fig. 8

Signaling pathways in bovine granulosa cells after DHA or TUG-891 treatment. Effects of DHA or TUG-891 on phosphorylation of (a) mitogen-activated protein kinase 14 (MAPK14), (b) AMP-activated protein kinaseα (AMPKα) and (c) protein kinase B (Akt) signaling pathways were assessed in bovine granulosa cells cultured for 15 h in enriched McCoy’s 5A media with 10 μM DHA or 1 μM TUG-891, as described in Material and Method section for 5, 10, 30 and 60 min. Protein extracts were separated by electrophoresis on 4–12% (w:v) SDS-polyacrylamide gel. After electrotransfer to nitrocellulose membranes, the proteins were probed with anti-phosphorylated (p-)MAPK14 (a), anti-p-AMPKα (b) or anti-p-AKT1/2/3 (c) antibodies. The blots were stripped and re-probed with antibodies against MAPK14, AMPKα or Akt, respectively. Bands on the blots were quantified. Results of four independent experiments are presented as the ratio of p-protein to total protein, normalized by the ratio observed in control at each time and expressed as mean ± SEM of four independent experiments, with time 0 min being equal to 1 (for reference). Bars with different superscripts are significantly different (p < 0.05)

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