Journal: Frontiers in Molecular Biosciences

Article Title: Expression and distribution of activin-follistatin-inhibin axis in the urinary bladder

doi: 10.3389/fmolb.2025.1519977

Figure Lengend Snippet: Antibody list.

Article Snippet: Inha , BIOSS , Bs-1032R , IF, 1:100.

Techniques:

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: Six-week-old female rats received OVX or a 0.25 mg E2 pellet was implanted subcutaneously after OVX (OVX + E2). Sham-operated (ovary-intact) rats were used as a control (A–C). In another series of experiments, a 0.25 mg E2 pellet was implanted subcutaneously in ovary-intact 6-week-old female rats. Sham-operated (no E2 pellet) rats were used as a control (D–F). Seven days later, the rats were euthanized, and the posterior part of the hypothalamus was removed. mRNA was extracted from the hypothalamic tissues and reverse transcribed. The mRNA levels of inhibin α (A and D), βA (B and E), and βB (C and F) were measured by quantitative RT-PCR. Samples for each experimental group were run in duplicate and normalized to the mRNA levels of GAPDH as a housekeeping gene. The results are expressed as fold induction over control and presented as the mean ± SEM. ** p < 0.01, * p < 0.05 vs. control.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Control, Reverse Transcription, Quantitative RT-PCR

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: (A) Total RNA was extracted from rHypoE8 and GT1-7 cells, and RT-PCR was carried out for 40 cycles using primers specific to Kiss1 , inhibin α, inhibin βA, and inhibin βB. PCR products were resolved in a 1.5% agarose gel and visualized with ethidium bromide staining. mRNA from rat brain tissues was used as a positive control. (B) Cell lysates (10 μg protein) from rHypoE8 and GT1-7 cells were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by immunoblotting and incubation with antibodies against kisspeptin, inhibin α, inhibin βA, and inhibin βB. β-Actin was detected as an internal control. Proteins from rat brain tissues were used as a positive control. The bands were visualized using a horseradish peroxidase-conjugated secondary antibody.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Positive Control, Polyacrylamide Gel Electrophoresis, Western Blot, Incubation, Control

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: rHypoE8 (A–C) and GT1-7 (D–F) cells were stimulated with 10 or 100 nM E2 for 24 h, after which mRNA was extracted and reverse transcribed. Inhibin α (A and D), βA (B and E), and βB (C and F) subunit mRNA levels were measured by quantitative RT-PCR. Results are expressed as fold induction over unstimulated cells and presented as mean ± SEM values of three independent experiments, each performed with duplicate samples. ** p < 0.01, * p < 0.05 vs. control.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Reverse Transcription, Quantitative RT-PCR, Control

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: rHypoE8 (A–C) and GT1-7 (D–F) hypothalamic cell models were stimulated with the indicated concentrations of activin (A and D), inhibin A (B and E), or inhibin B (C and F) for 24 h. After which, mRNA was extracted and reverse transcribed. Kiss1 mRNA levels were measured by quantitative RT-PCR. Results are expressed as fold induction over unstimulated cells and presented as mean ± SEM values of three independent experiments, each performed with duplicate samples. ** p < 0.01, * p < 0.05 vs. control.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Reverse Transcription, Quantitative RT-PCR, Control

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: rHypoE8 (A–C) and GT1-7 (D–F) hypothalamic cell models were stimulated with the indicated concentrations of KP10 for 24 h. After which, mRNA was extracted and reverse transcribed. Inhibin α (A and D), βA (B and E), and βB (C and F) subunit mRNA levels were measured by quantitative RT-PCR. Results are expressed as fold induction over unstimulated cells and presented as mean ± SEM values of three independent experiments, each performed with duplicate samples. ** p < 0.01, * p < 0.05 vs. control.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Reverse Transcription, Quantitative RT-PCR, Control

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: mHypoA55 cells were stimulated with the indicated concentrations of KP10 for 24 h. After which, mRNA was extracted and reverse transcribed. Inhibin α (A), βA (B), and βB (C) subunit mRNA levels were measured by quantitative RT-PCR. Results are expressed as fold induction over unstimulated cells and presented as mean ± SEM values of three independent experiments, each performed with duplicate samples. * p < 0.05 vs. control.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Reverse Transcription, Quantitative RT-PCR, Control

Journal: Endocrine Journal

Article Title: Potential role of kisspeptin in the estradiol-induced modulation of inhibin subunit gene expression: Insights from in vivo rat models and hypothalamic cell models

doi: 10.1507/endocrj.EJ25-0044

Figure Lengend Snippet: Inhibin α subunit expression in the posterior area of the hypothalamus was increased following OVX, and this increase was suppressed by E2 supplementation. This pattern mirrors the well-established E2-mediated suppression of Kiss1 gene expression in this region. Because kisspeptin stimulation significantly increased inhibin α subunit expression, and because neither activin nor inhibin modulated Kiss1 gene expression in a variety of hypothalamic cells, we speculate that kisspeptin may act as an upstream regulator of inhibin and activin expression in the hypothalamus.

Article Snippet: Protein was transferred onto polyvinylidene difluoride membranes (Hybond-P PVDF; Amersham Biosciences, Little Chalfont, UK), which were blocked for 1 h at room temperature in Blotto (5% milk in Tris-buffered saline), including an anti-kisspeptin antibody (1:100 dilution; Abcam) [ ], anti-inhibin α antibody (1:100 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX) [ ], anti-inhibin βA antibody (1:100 dilution; Santa Cruz Biotechnology, Inc.) [ ], anti-inhibin βB antibody (1:1,000 dilution; Abcam) [ ], or anti-β-actin primary antibody (1:1,000 dilution; Abcam) [ ] in Blotto overnight at 4°C and washed three times for 5 min per wash with Tris-buffered saline/1% Tween.

Techniques: Expressing, Gene Expression