Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Effects of Levonorgestrel and progesterone on Oviductal physiology in mammals

doi: 10.1186/s12958-018-0377-3

Figure Lengend Snippet: Primer sequences of PCR

Article Snippet: Antibodies against β-actin (1:5000, Proteintech, IL, USA), IGFBP1 (1:1000, Proteintech, IL, USA), ITGB3 (1:1000, Proteintech, IL, USA), MUC1 (1:1000, Proteintech, IL, USA), ACVR1B (1:1000, Proteintech, IL, USA) and STAT3 (1:1000, Cell Signaling Technology, Danvers, MA, USA) were used as primary antibodies, and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:5000, Cell Signaling Technology, Danvers, MA, USA) and goat anti-mouse IgG (1:5000, Cell Signaling Technology, Danvers, MA, USA) were used as secondary antibodies.

Techniques:

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Effects of Levonorgestrel and progesterone on Oviductal physiology in mammals

doi: 10.1186/s12958-018-0377-3

Figure Lengend Snippet: Effects of different concentrations of LNG and P4 on the expression of receptivity markers in the fallopian tubes. a-b mRNA expression levels of LIF, STAT3, IGFBP1, ITGB3, MUC1, and ACVR1B in OE-E6/E7 cells following treatment with different concentrations of LNG and P4 ( n = 3 in each group; ns, not significant); c protein expression levels of MUC1, ITGB3, ACVR1B, STAT3, and IGFBP1 in OE-E6/E7 cells following treatment with different concentrations of LNG and P4

Article Snippet: Antibodies against β-actin (1:5000, Proteintech, IL, USA), IGFBP1 (1:1000, Proteintech, IL, USA), ITGB3 (1:1000, Proteintech, IL, USA), MUC1 (1:1000, Proteintech, IL, USA), ACVR1B (1:1000, Proteintech, IL, USA) and STAT3 (1:1000, Cell Signaling Technology, Danvers, MA, USA) were used as primary antibodies, and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:5000, Cell Signaling Technology, Danvers, MA, USA) and goat anti-mouse IgG (1:5000, Cell Signaling Technology, Danvers, MA, USA) were used as secondary antibodies.

Techniques: Expressing

Journal: bioRxiv

Article Title: Sox9 links biliary maturation to branching morphogenesis

doi: 10.1101/2024.01.15.574730

Figure Lengend Snippet: (A) Bulk RNA-seq identifies DEGs between Sox9cKO and control BECs. (B) GSEA demonstrates significant enrichment of TGF-β signaling in Sox9cKO BECs relative to control BECs. (C) Co-localization of pSMAD2 and EpCAM confirms elevated TGF-β signaling in Sox9cKO BECs (scale bar represents 5μm; white asterisks indicate portal vein). (D) IF demonstrates elevated Activin A in EpCAM+ BECs in Sox9cKO livers (scale bar represents 50μm; white asterisks indicate portal vein). (E) GSEA demonstrates that Sox9cKO BECs are de-enriched for adult BEC gene signature (NES: −2.70; p = 3.67E-10) and enriched for adult hepatocyte gene signature (NES: 2.31; p = 3.67E-10). (F) GSEA of liver development gene sets shows that the Sox9cKO BEC transcriptome resembles embryonic BECs (NES: 1.23; p = 6.06E-6) and embryonic hepatocytes (NES: 2.32; p = 3.00E-10), but not hepatoblasts (NES: 0.99; p = 0.55). (G) Co-localization of NCAM1 and EpCAM is significantly elevated in Sox9cKO compared to controls (scale bar represents 5μm; white asterisks indicate portal vein). (H) scRNA-seq of FACS-isolated EpCAM+ BECs reveals subpopulation-specific transcriptomic changes in Sox9cKO. (I) Upregulation of Inhba and Ncam1 in Sox9cKO samples is specific to Sox9cKO ductule-like cells (cluster 6).

Article Snippet: To neutralize Activin A, control and Sox9cKO neonates were administered 2 µg/g anti-Activin A (Biotechne; MAB3381) or mouse IgG 1 (Biotechne; MAB002) at P1 and P3 by intraperitoneal injection.

Techniques: RNA Sequencing Assay, Isolation

Journal: bioRxiv

Article Title: Sox9 links biliary maturation to branching morphogenesis

doi: 10.1101/2024.01.15.574730

Figure Lengend Snippet: (A) Single Sox9cKO BECs cultured for 7d fail to form lumens typical of control mICOs. (B) Sox9cKO BECs form significantly fewer mICOs, a majority of which (C) exhibit non-cystic morphology. (D) mICO area is unchanged between control and Sox9cKO. (E) 50ng/mL Activin A is sufficient to induce Sox9cKO morphology in WT BECs, but (F) unlike Sox9cKO (Sox9 Δ/Δ) did not impact mICO organoid formation. (G) Rates of non-cystic mICO morphology were similar between Sox9cKO and Activin A treated WT BECs. (H) Organoid area remained unchanged between all groups. (I) Ncam1 was significantly upregulated in Sox9cKO mICOs by RT-qPCR, and induced by Activin A in WT mICOs. (J) Activin A upregulates Sox9 expression in WT mICOs (letters indicate grouping by significance , p < 0.05; scale bars represent 50μm).

Article Snippet: To neutralize Activin A, control and Sox9cKO neonates were administered 2 µg/g anti-Activin A (Biotechne; MAB3381) or mouse IgG 1 (Biotechne; MAB002) at P1 and P3 by intraperitoneal injection.

Techniques: Cell Culture, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Sox9 links biliary maturation to branching morphogenesis

doi: 10.1101/2024.01.15.574730

Figure Lengend Snippet: (A) Co-localization of Ki-67 and EpCAM in IgG and α-ActA treated livers demonstrates that Activin A neutralization in vivo (B) increases BEC numbers per portal field while (C) inhibiting BEC proliferation (scale bar represents 50μm; white asterisks indicate portal vein). (D) Whole tissue imaging of EpCAM+ IHBDs reveals that Activin A neutralization partially rescues Sox9cKO ductal paucity, resulting in increased branching near the hilum. (E) Sholl analysis supports increased proximal branching and decreased distal branching in α-ActA treated livers, which is corroborated by (F) AUC demonstrating increased hilar branch complexity (0.00-0.25a.u.), no change at 0.25-0.50a.u., and decreased branching distally (0.50-1.00a.u.). (G) Sholl decay coefficient is increased following Activin A neutralization reflecting increased localized branching near the hilum.

Article Snippet: To neutralize Activin A, control and Sox9cKO neonates were administered 2 µg/g anti-Activin A (Biotechne; MAB3381) or mouse IgG 1 (Biotechne; MAB002) at P1 and P3 by intraperitoneal injection.

Techniques: Neutralization, In Vivo, Imaging

Journal: bioRxiv

Article Title: A novel dephosphorylation peptide inhibits 17β-HSD1 enzyme activity in ovarian granulosa cells and breast cancer cells

doi: 10.64898/2025.12.29.696806

Figure Lengend Snippet: Activin A and IGF-1 can facilitate 17β-HSD1 phosphorylation and E2 synthesis A. After treating porcine granulosa cells with activin A for different time periods, Western blot was used to detect the expression of 17β-HSD1, phosphorylation at key sites, and the total phosphorylation level. The experiment was repeated five times. B-F. Gray value analysis of the expression of 17β-HSD1, phosphorylation at key sites, and the total phosphorylation level. Different superscript letters indicate significant differences between groups. G. Changes in the concentration of E2 in the cell culture medium of porcine granulosa cells after treating with activin A for 24 h. H. Detection of the protein expression of 17β-HSD1, 17β-HSD1 Ser30, 17β-HSD1 Ser274, and total phosphorylation of 17β-HSD1 in MCF-7 cells treated with activin A (50 ng/mL) using Western blotting; I. Analysis of the relative expression levels of 17β-HSD1 and its phosphorylation by densitometry; J. Representative pictures of three-dimensional culture of mouse follicles, bar=100 μm. K. Comparison of follicle survival rates after activin A treatment. L. Comparison of follicle growth diameters after activin A treatment. M. After injecting 80 μg IGF-1 into KM mice for 24 hours, Western blot was used to detect the expression of 17β-HSD1 and the phosphorylation level at key sites in the ovary. N. Quantitative analysis of the gray value in M. O. Blood was collected at different time points after injecting 80 μg IGF-1 into KM mice, and the concentration of serum E2 was detected. Five mice were repeated at each time point. The “*” indicates a significant difference between groups.

Article Snippet: Activin A were purchased from MCE (HY-P70311), 50 ng/mL activin A were used for cell treatment and follicle cultuer.

Techniques: Phospho-proteomics, Western Blot, Expressing, Concentration Assay, Cell Culture, Comparison

Journal: bioRxiv

Article Title: A novel dephosphorylation peptide inhibits 17β-HSD1 enzyme activity in ovarian granulosa cells and breast cancer cells

doi: 10.64898/2025.12.29.696806

Figure Lengend Snippet: The treatment of SS-CPP reduces the phosphorylation level of 17β-HSD1 A. The effects of porcine granulosa cells treated with and without activin A and different CPPs on the phosphorylation at the key sites and total phosphorylation of 17β-HSD1 were investigated. B. The relative quantitative analysis of the expression of 17β-HSD1 Ser30 site relative to 17β-HSD1. C. The relative quantitative analysis of the expression of 17β-HSD1 Ser274 site relative to 17β-HSD1. D. The relative quantitative analysis of the total phosphorylation level of 17β-HSD1 relative to 17β-HSD1 expression. E. Detection of changes in 17β-HSD1 and its phosphorylation levels in MCF-7 cells after co-treatment with cell-penetrating peptides and activin A for 12 hours using Western blotting. F. Analysis of relative expression levels of 17β-HSD1 by densitometry. G. Analysis of phosphorylation levels of 17β-HSD1 at Ser30 relative to 17β-HSD1 expression by densitometry. H. Analysis of phosphorylation levels of 17β-HSD1 at Ser274 relative to 17β-HSD1 expression by densitometry. I. Analysis of total phosphorylation levels of 17β-HSD1 relative to 17β-HSD1 expression by densitometry. Different superscript letters indicate significant differences between groups.

Article Snippet: Activin A were purchased from MCE (HY-P70311), 50 ng/mL activin A were used for cell treatment and follicle cultuer.

Techniques: Phospho-proteomics, Expressing, Western Blot

Journal: bioRxiv

Article Title: A novel dephosphorylation peptide inhibits 17β-HSD1 enzyme activity in ovarian granulosa cells and breast cancer cells

doi: 10.64898/2025.12.29.696806

Figure Lengend Snippet: SS-CPP inhibited the growth of tumors in MCF-7 breast cancer-bearing nude mice A. Nude mice and the dissected tumor masses on the 31st day after tumor formation. Starting from the 21st day after tumor formation, the control group was injected with 0.1 mL of normal saline daily, the activin A group was injected with 0.1 mL of 100 ng/0.1 mL activin A daily, and the SS-CPP group was injected with 0.1 mL of SS-CPP synthetic polypeptide (combination of 30S-CPP and 274S-CPP) at a concentration of 75 µg/0.1 mL daily. B. Comparison of tumor weights on the 31st day after tumor formation. C-E. Growth curves of tumor masses, including the changes in tumor diameter, length, and width during the experiment. F. The change curve of the body weight of nude mice during the experiment. G. Effect of SS-CPP treatment on the proliferation and apoptosis of the formed tumor tissue. Ki67 was labeled with red fluorescence, and cleaved caspase 3 was labeled with green fluorescence. The “*” indicates a significant difference between groups.

Article Snippet: Activin A were purchased from MCE (HY-P70311), 50 ng/mL activin A were used for cell treatment and follicle cultuer.

Techniques: Control, Injection, Saline, Concentration Assay, Comparison, Labeling, Fluorescence