Fig 1: The mRNA expression of PPARA (A), PPARD (B) and PPARG (C) in the bovine CL explants after 24 h of incubation with PGF2a (10-6 M) on days 15–17 of the estrous cycle. The mRNA expression profiles are presented as a fold change. Presented results are the mean ± SEM from 9 animals. The asterisks indicate statistical differences (* p < 0.05, ** p < 0.01) between control (C) and PGF2a-treated (P) explants, as determined by the nonparametric Mann–Whitney U test.
Fig 2: The mRNA expression of PPARA (A), PPARD (B) and PPARG (C), and tissue concentration of PPARa (D), PPARd (E) and PPAR? (F) in the bovine CL on days 2–3, 5–6, 8–12, 15–17 and 19–21 of the estrous cycle. The mRNA expression profiles are presented in arbitrary units as the ratio of expression of the target genes to the mean of the best combination of two reference genes, namely, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ribosomal 18S RNA (RN18S1). The concentration is expressed as pg/g tissue. Presented results are the mean ± SEM from 8 animals. The superscript letters “a, b, c” indicate the statistical differences between groups, as determined by nonparametric one-way ANOVA Kruskal–Wallis followed by Dunn’s multiple comparisons test.
Fig 3: Scheme 1 of the estrous cycle (n = 9). The CL explants were preincubated in vitro for 2 h. Next, explants were cultured in the presence of PPAR antagonists: PPARa antagonist (10-5 M, GW6471; Sigma-Aldrich, Saint Louis, MO, USA, G5045), PPARd antagonist (10-5 M, GSK3787; Sigma-Aldrich, Saint Louis, MO, USA, G7423) and PPAR? antagonist (10-5 M, GW9662; Cayman Chemical, Ann Arbor, MI, USA, 70785), in combination with or without PGF2a receptor (FP) antagonist (10-5 M, AL8810; Sigma-Aldrich, Saint Louis, MO, USA, A3846) to block receptor action for 6 h, and then CL explants were stimulated with PGF2a (10-6 M; Sigma-Aldrich, Saint Louis, MO, USA, P5069) for a further 24 h without medium exchange. Experimental groups were marked as follows: C—control group (untreated CL explants); P—CL explants stimulated with PGF2a; PAL—CL explants stimulated with FP antagonist and PGF2a; APAL 1/2—CL explants stimulated with FP antagonist, PPARa antagonist, PPARd antagonist and PGF2a; APAL 2/3—CL explants stimulated with FP antagonist, PPARd antagonist, PPAR? antagonist and PGF2a; APAL 1/3—CL explants stimulated with FP antagonist, PPARa antagonist, PPAR? antagonist and PGF2a; and AP 1/2/3—CL explants stimulated with PPARa antagonist, PPARd antagonist, PPAR? antagonist and PGF2a. After incubation, the culture medium was frozen at -20 °C until further determination using RIA. Tissue explants were frozen at -80 °C until further analysis using qPCR.
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