Journal: Molecular Medicine Reports

Article Title: Uremic toxin p-cresyl sulfate enhances the calcification of aortic valvular interstitial cells via klotho/sirtuin-1 signaling

doi: 10.3892/mmr.2026.13872

Figure Lengend Snippet: Effect of PCS on klotho and HIF-1α expression. Western blotting and semi-quantification of (A) klotho and (B) HIF-1α protein expression in control and PCS-treated valvular interstitial cells. PCS-treated cells were incubated with PCS (100 µM) for 7 days (n=5). β-actin was used as an internal control. *P<0.05. PCS, p-cresol; HIF-1α, hypoxia-inducible factor-1α.

Article Snippet: The membranes were blocked with 10% non-fat milk in PBS for 1 h at room temperature and incubated overnight at 4°C with primary antibodies against RUNX2 (1:2,000 dilution; cat. no. 12556; Cell Signaling Technology), acetylated-NF-κB (1:2,000 dilution; acetyl-NF-κB; cat. no. 3045; Cell Signaling Technology, Inc.), NF-κB (1:3,000 dilution; cat. no. 8242; Cell Signaling Technology, Inc.), HIF-1α (1:1,000 dilution; cat. no. NB100-479; Novus Biologicals), klotho (1:1,000 dilution; cat. no. LS-C145689; LifeSpan BioSciences, Inc.) and β-actin (1:6,600 dilution; cat. no. ab6276; Abcam).

Techniques: Expressing, Western Blot, Control, Incubation

Journal: Molecular Medicine Reports

Article Title: Uremic toxin p-cresyl sulfate enhances the calcification of aortic valvular interstitial cells via klotho/sirtuin-1 signaling

doi: 10.3892/mmr.2026.13872

Figure Lengend Snippet: Effect of HIF-1α inhibition on PCS-induced VIC calcification. The upper panel depicts representative images of Alizarin Red S-stained VICs that have been incubated with or without the HIF-1α inhibitor PX-478 (0.5 µM) and with or without PCS (100 µM). VIC calcification was quantified by determining the total area of positive staining (red) per field (3 fields were observed per treatment group), as shown in the lower panel (n=3). Scale bar, 25 µm. *P<0.05. PCS, p-cresol; VIC, valvular interstitial cell; HIF-1α, hypoxia-inducible factor-1α.

Article Snippet: The membranes were blocked with 10% non-fat milk in PBS for 1 h at room temperature and incubated overnight at 4°C with primary antibodies against RUNX2 (1:2,000 dilution; cat. no. 12556; Cell Signaling Technology), acetylated-NF-κB (1:2,000 dilution; acetyl-NF-κB; cat. no. 3045; Cell Signaling Technology, Inc.), NF-κB (1:3,000 dilution; cat. no. 8242; Cell Signaling Technology, Inc.), HIF-1α (1:1,000 dilution; cat. no. NB100-479; Novus Biologicals), klotho (1:1,000 dilution; cat. no. LS-C145689; LifeSpan BioSciences, Inc.) and β-actin (1:6,600 dilution; cat. no. ab6276; Abcam).

Techniques: Inhibition, Staining, Incubation

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: Expression and localization of key proteins in the PI3K-AKT/HIF-1α signaling pathway in normal yak testes.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: Expressing

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: Effects of hypoxia on testicular development in mice. ( A ) Histological analysis of mouse testes before and after hypoxia treatment. ( B ) Testicular changes in mice after 14 days of hypoxia treatment. ( C ) Changes in HIF-1α contents in the blood of mice in the control and hypoxia groups. ( D ) The expression levels of genes and proteins in hypoxia-related pathways in the testes of mice in the control and hypoxia groups. ( E ) Gel map of proteins in hypoxia-related pathways in the testes of mice in the control and hypoxia groups. Data are presented as mean ± SEM. ** p < 0.01; * p < 0.05.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: Control, Expressing

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: Analysis of the expression of key proteins in the PI3K-AKT/HIF-1α signaling pathway in mouse testes. ( A ) Immunofluorescence images of key proteins of PI3K-AKT/HIF-1α signaling pathway and CAMs in mouse testes after hypoxia treatment; ( B ) Bar chart of quantitative analysis of fluorescence intensity. Data are presented as mean ± SEM. ** p < 0.01; * p < 0.05; ns (not significant), p > 0.05.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: Expressing, Immunofluorescence, Fluorescence

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: The effect of in vitro injection of inhibitors on the expression of adhesion molecules in mouse testicular cells. ( A ) The relative mRNA expression of key proteins in the PI3K-AKT/HIF-1α signaling pathway and CAMs after inhibitor treatment. ( B ) Western blot gels of key protein expression levels in the PI3K-AKT/HIF-1α signaling pathway and CAMs following inhibitor treatment. ( C ) After treatment with different inhibitors in mice, the protein expression levels of key molecules in the PI3K-AKT/HIF-1α signaling pathway and CAMs were plotted. Data are presented as mean ± SEM. ★, p < 0.01; ☆, p < 0.05; #, p > 0.05.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: In Vitro, Injection, Expressing, Western Blot

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: The expression of key proteins in the PI3K-AKT/HIF-1α signaling pathway and CAMs in mouse Sertoli cells. ( A ) Identification of mouse testicular Sertoli cells. ( B ) The expression of key proteins in the PI3K-AKT/HIF-1α signaling pathway in mouse Sertoli cells. ( C ) Expression of CAMs in mouse Sertoli cells.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: Expressing

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: The effect of inhibiting the expression of key proteins in the PI3K-AKT/HIF-1α signaling pathway on the expression of adhesion molecules in mouse Sertoli cells. ( A ) Mouse Sertoli cell administration pattern diagram. ( B ) Determination of the optimal concentration of the two inhibitors BAY 87-2243 and LY294002. ( C – J ) The relative mRNA expression of key proteins in the PI3K-AKT/HIF-1α signaling pathway and CAMs after inhibitor treatment. ( K ) Western blot gels of key protein expression levels in the PI3K-AKT/HIF-1α signaling pathway and CAMs following inhibitor treatment. ( L – R ) After treatment of Sertoli cells with the different inhibitors, the protein expression levels of key molecules in the PI3K-AKT/HIF-1α signaling pathway and CAMs were plotted. Data are presented as mean ± SEM. ** p < 0.01; * p < 0.05; ns (not significant), p > 0.05.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: Expressing, Concentration Assay, Western Blot

Journal: Veterinary Sciences

Article Title: The HIF-Mediated PI3K-AKT Signaling Pathway Is a Key Signaling Pathway Triggering Testicular Spermatogenic Disorders in Yaks with Cryptorchidism

doi: 10.3390/vetsci13050468

Figure Lengend Snippet: Schematic diagram showing how HIF-1α regulates the adhesion mechanism of mouse testicular supporting cells via the PI3K-AKT signaling pathway. Graphical abstract: Under hypoxia, blood oxygen content is decreased, which increases the protein concentration of HIF-1α; this, in turn, stimulates PI3K-AKT signaling pathway-mediated synthesis of Sertoli CAMs. Impaired synthesis of testicular adhesion molecules directly affects testicular spermatogenesis and can even induce a series of male reproductive system diseases, such as testicular atrophy or cryptorchidism.

Article Snippet: HIF-1α , Affinity Biosciences, USA , BF8002 , WB:1:400 IF/IH:1:500.

Techniques: Protein Concentration