rabbit anti-foxl2 purified antibody  (CASLO Inc)

Journal: International Journal of Molecular Sciences

Article Title: FOXL2 is a Progesterone Target Gene in the Endometrium of Ruminants

doi: 10.3390/ijms21041478

Figure Lengend Snippet: FOXL2 expression in ovine endometrium. Caruncular (CAR) and intercaruncular (ICAR) endometrial areas were collected from cyclic Pré-alpes du sud ewes at various days post- oestrus (day 4, n = 4; day 8, n = 4; day 12, n = 4). Two experimental groups were subsequently added to this study, then CAR and ICAR endometrial areas were collected from cyclic ( n = 4) and pregnant ( n = 4) Pré-alpes du sud ewes at 15 days post- oestrus . ( A ) Quantification of FOXL2 mRNA by RT-qPCR. Expression of FOXL2 gene was normalised against that of RPL19 . ( B ) Quantification of FOXL2 protein by Western blotting. The amount of FOXL2 was normalized to that of ACTB Quantitative data are presented as the mean ± SEM. * p < 0.05.

Article Snippet: Total proteins were extracted from frozen tissue and Western blot immunoassays were processed with 15 μg of total protein extract, as described previously [ ], A rabbit anti-FOXL2 purified antibody generated against a peptide corresponding to the C-terminal conserved region of mammalian FOXL2 (WDHDSKTGALHSRLDL, diluted 1:500, CASLO Laboratory, Denmark) was used in PBS-T solution containing 4% non-fat dry milk and then, a goat peroxidase-conjugated anti-rabbit IgG antibody (diluted 1:5000, SantaCruz Biotechnology; Heidelberg, Germany).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: FOXL2 is a Progesterone Target Gene in the Endometrium of Ruminants

doi: 10.3390/ijms21041478

Figure Lengend Snippet: Regulation of FOXL2 gene expression in ewes with lower circulating P4 concentrations. Caruncular (CAR) and intercaruncular (ICAR) endometrial areas were collected from pregnant Pré-alpes du sud ewes treated with DMSO as a control solution ( n = 7) or with trilostane (15 mg/ewe in 1 mL DMSO; twice a day, n = 10) for 11 days. ( A ) Progesterone dosage throughout the treatment: DMSO ( n = 7) or Trilostane ( n = 10) for 11 days. ( B ) Quantification of FOXL2 mRNA by RT-qPCR. Expression of FOXL2 was normalized to that of RPL19. ( C ) Quantification of FOXL2 protein by Western blotting. The amount of FOXL2 was normalized to that of ACTB. Quantitative data are presented as mean ± SEM. * p < 0.05.

Article Snippet: Total proteins were extracted from frozen tissue and Western blot immunoassays were processed with 15 μg of total protein extract, as described previously [ ], A rabbit anti-FOXL2 purified antibody generated against a peptide corresponding to the C-terminal conserved region of mammalian FOXL2 (WDHDSKTGALHSRLDL, diluted 1:500, CASLO Laboratory, Denmark) was used in PBS-T solution containing 4% non-fat dry milk and then, a goat peroxidase-conjugated anti-rabbit IgG antibody (diluted 1:5000, SantaCruz Biotechnology; Heidelberg, Germany).

Techniques: Gene Expression, Control, Quantitative RT-PCR, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: FOXL2 is a Progesterone Target Gene in the Endometrium of Ruminants

doi: 10.3390/ijms21041478

Figure Lengend Snippet: FOXL2 endometrial expression in ovariectomized ewes supplemented with ovarian steroids. Caruncular (CAR) and intercaruncular (ICAR) endometrial areas were collected from ovariectomized Pré-alpes du sud ewes supplemented with a control solution (OVX, n = 4), (P4) solution (OVX + P4, n = 4), (E2) solution (OVX + E2, n = 4) or (P4 + E2) solution (OVX + E2 + P4, n = 4) for 12 days and also from cyclic ewes at 12 days ( n = 4). ( A ) Quantification of FOXL2 mRNA by RT-qPCR. Expression of FOXL2 was normalized to that of RPL19. ( B ) Quantification of FOXL2 protein by Western blotting. The amount of FOXL2 was normalized to that of ACTB. Quantitative data are presented as mean ± SEM. * p < 0.05.

Article Snippet: Total proteins were extracted from frozen tissue and Western blot immunoassays were processed with 15 μg of total protein extract, as described previously [ ], A rabbit anti-FOXL2 purified antibody generated against a peptide corresponding to the C-terminal conserved region of mammalian FOXL2 (WDHDSKTGALHSRLDL, diluted 1:500, CASLO Laboratory, Denmark) was used in PBS-T solution containing 4% non-fat dry milk and then, a goat peroxidase-conjugated anti-rabbit IgG antibody (diluted 1:5000, SantaCruz Biotechnology; Heidelberg, Germany).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: FOXL2 is a Progesterone Target Gene in the Endometrium of Ruminants

doi: 10.3390/ijms21041478

Figure Lengend Snippet: FOXL2 endometrial expression under the influence of ovarian steroid hormones balance in ovariectomized cows and bovine explants. The expression of FOXL2 was quantified by RT-qPCR, and normalized to RPL19. ( A ) Strips of endometrial tissue were collected from ovariectomized cows supplemented with a control solution (OVX; n = 3), progesterone (OVX + P4; n = 3), oestradiol (OVX + E2; n = 3) or both steroids (OVX + E2 + P4; n = 3). Data were analysed by ANOVA and are presented as the mean ± SEM. Bars with different superscripts significantly differ ( p < 0.05). ( B ) Intercaruncular endometrial explants from two cows were cultured ex vivo for 48 h in control medium (C), or medium containing 5 ng/mL progesterone (P4) or 3 pg/mL oestradiol (E2). The expression of FOXL2 and PGR transcripts was quantified by RT-qPCR and normalized to RPL19 gene expression. Data were analysed by ANOVA and are presented as the mean ± SEM. * p < 0.05.

Article Snippet: Total proteins were extracted from frozen tissue and Western blot immunoassays were processed with 15 μg of total protein extract, as described previously [ ], A rabbit anti-FOXL2 purified antibody generated against a peptide corresponding to the C-terminal conserved region of mammalian FOXL2 (WDHDSKTGALHSRLDL, diluted 1:500, CASLO Laboratory, Denmark) was used in PBS-T solution containing 4% non-fat dry milk and then, a goat peroxidase-conjugated anti-rabbit IgG antibody (diluted 1:5000, SantaCruz Biotechnology; Heidelberg, Germany).

Techniques: Expressing, Quantitative RT-PCR, Control, Cell Culture, Ex Vivo, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: FOXL2 is a Progesterone Target Gene in the Endometrium of Ruminants

doi: 10.3390/ijms21041478

Figure Lengend Snippet: Progesterone regulates FOXL2 promoter activity in vitro. COS7 cells were cultivated for few passages then transfected using Xtreme gene transfection reagent with progesterone receptor (PGR) A and/or B expressing vectors as well as FOXL2-promoter sequence (1 kb) associated with the luciferase gene for 48 h in the presence or absence of a progesterone (P4) treatment. Activity of the FOXL2 promoter was normalized to TK–Renilia vector activity. Quantitative data are presented as the mean ± SEM. ** p < 0.01; *** p < 0.001.

Article Snippet: Total proteins were extracted from frozen tissue and Western blot immunoassays were processed with 15 μg of total protein extract, as described previously [ ], A rabbit anti-FOXL2 purified antibody generated against a peptide corresponding to the C-terminal conserved region of mammalian FOXL2 (WDHDSKTGALHSRLDL, diluted 1:500, CASLO Laboratory, Denmark) was used in PBS-T solution containing 4% non-fat dry milk and then, a goat peroxidase-conjugated anti-rabbit IgG antibody (diluted 1:5000, SantaCruz Biotechnology; Heidelberg, Germany).

Techniques: Activity Assay, In Vitro, Transfection, Expressing, Sequencing, Luciferase, Plasmid Preparation