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

Journal: Pharmaceuticals

Article Title: Polyoxypregnane Aryl Esters Prepared from Metaplexis japonica (Thunb.) Makino and Their Role in Reversing Multidrug Resistance in HepG2/Dox Cells

doi: 10.3390/ph18081187

Figure Lengend Snippet: Response (RU)–concentration curves from SPR analyses of the binding of compound 1 or 1a to full-length human ABCB1.

Article Snippet: The human recombinant ABCB1 full-length protein (HEK293, Flag) (MedChemExpress, Monmouth Junction, NJ, USA; #HY-P703175) was coupled onto a CM5 sensor chip (Cytiva, Marlborough, MA, USA; #BR-1005-30) via carboxyl groups on the dextran.

Techniques: Concentration Assay, Binding Assay

Journal: Frontiers in pharmacology

Article Title: Follistatin Attenuates Myocardial Fibrosis in Diabetic Cardiomyopathy via the TGF-β-Smad3 Pathway.

doi: 10.3389/fphar.2021.683335

Figure Lengend Snippet: FIGURE 5 | Overexpression of FST reduces cardiac fibrosis by inhibiting the activation of the TGF-β–Smad3 pathway in vivo and in vitro. Quantitative real-time PCR analysis of TGF-β1, activin A, and TGF-β3 mRNA transcription [(A–C), n≥4] and p-Smad3 and Smad3 protein expression in each group [(D), n 4]. In vitro, NIH3T3 cells were treated with or without high glucose (30 mM), recombinant FST protein (10 ng/ml), and TGF-β1 (20 μM). Protein expression of COL1, COL3, p-Smad3, and Smad3 with different treatments [(E–G), n 3]. *p< 0.05; **p< 0.01; ***p<0.001.

Article Snippet: After starvation in serum-free medium for 24 h, the NIH3T3 cells were incubated in DMEM containing 5.5 mmol/L glucose, 33 mmol/L glucose (high glucose, HG), HG plus 10 ng/ml recombinant FST protein (human recombinant follistatin, BioVision, #4708), or HG plus 10 ng/ml recombinant FST protein and 20 μM TGF-β1 (MedChemExpress, United States).

Techniques: Over Expression, Activation Assay, In Vivo, In Vitro, Real-time Polymerase Chain Reaction, Expressing, Recombinant