horseradish peroxidase conjugated goat anti rabbit secondary antibody  (Cell Signaling Technology Inc)

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: Notch2 overexpression enhanced ovarian functions in premature ovarian failure (POF) mice by activating the Wnt/β-catenin pathway. ( A ) Quantitative real-time PCR (qPCR) for the validation of oe-Notch2 overexpression efficiency. ( B ) Representative images of ovarian size at the end of the study. ( C ) Quantification of ovarian weight at the end of the study. ( D ) Enzyme-linked immunosorbent assay (ELISA) for follicle-stimulating hormone (FSH), estradiol (E2) and anti-mullerian hormone (AMH) levels in serum. ( E ) qPCR for the expressions of steroidogenic enzyme-related genes (StARD1, Cyp11a1 and 3-β-HSD) in ovarian tissues. ( F ) Representative images of hematoxylin-eosin (HE) staining and quantification of primordial follicles, primary follicles, secondary follicles, antral follicles, atretic follicles and mature follicles for ovarian tissues. Magnification×200, scale bar = 100 μm; Magnification×40, scale bar = 500 μm. PF: primitive follicle; PrF: primary follicle; SF: secondary follicle; AnF: antral follicle; AF: atretic follicle; MF: mature follicle. △ P < 0.05 and △△ P < 0.01 vs. Normal; P < 0.05 and P < 0.01 vs. Control; # P < 0.05 and ## P < 0.01 vs. oe-NC; @ P < 0.05 and @@ P < 0.01 vs. oe-Notch2. Results were presented as mean ± standard deviation (SD)

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Staining, Control, Standard Deviation

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: Notch2 overexpression suppressed oxidative stress and ovarian cell apoptosis in POF mice by activating the Wnt/β-catenin pathway. ( A ) Levels of oxidative stress markers (malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD)) in ovarian tissues. ( B ) Flow cytometry for cell apoptosis in ovarian tissues. ( C ) Western blot analysis was applied to measure the cleaved caspase-3/caspase-3 ratio in ovarian tissues. ( D ) qPCR for the expressions of cell cycle-related genes (CDK4, CCND2 and P21) and apoptosis-related genes (Bax and Bcl2) in ovarian tissues. P < 0.05 and P < 0.01 vs. Control; # P < 0.05 and ## P < 0.01 vs. oe-NC; @ P < 0.05 and @@ P < 0.01 vs. oe-Notch2. Results were presented as mean ± SD

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Over Expression, Flow Cytometry, Western Blot, Control

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: Notch2 interacted with Wnt2 and β-catenin in POF mice. ( A ) qPCR was used to detect Wnt-2 and β-catenin mRNA expressions in the ovarian tissues. ( B ) Immunohistochemistry was applied to measure Notch2 protein expression in the ovarian tissues. Magnification×200, scale bar = 100 μm. ( C ) Western blot analysis was utilized to test Notch2 pathway-related protein expressions (Notch2, Jag1 and Hes2) and Wnt2/β-catenin pathway-related protein expressions (Wnt2, β-catenin, Axin2 and LEF1) in the ovaries. ( D ) Co-immunoprecipitation (Co-IP) was conducted to further verify the protein-protein interactions between Notch2 and Wnt2. P < 0.05 and P < 0.01 vs. Control; # P < 0.05 and ## P < 0.01 vs. oe-NC; @ P < 0.05 and @@ P < 0.01 vs. oe-Notch2. Results were presented as mean ± SD

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Immunohistochemistry, Expressing, Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Protein-Protein interactions, Control

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: Notch2 knockdown worsened GC functions in POF models by suppressed Wnt/β-catenin pathway ( A ) qPCR revealed the knockdown efficiency of sh-Notch2 for KNG cells. ( B ) Cell counting kit-8 (CCK-8) assay revealed cell viability for KNG cells. ( C ) Flow cytometry displayed cell cycle distribution for KNG cells. ( D ) Flow cytometry displayed cell apoptosis for KNG cells. ( E ) Western blot analysis displayed the cleaved caspase-3/caspase-3 ratio for KNG cells. ( F ) qPCR for the expressions of CDK4, CCND2, P21, Bax and Bcl2 mRNAs for KNG cells. ( G ) ELISA exhibited FSH, E2 and AMH levels for KNG cells. ( H ) qPCR for the expressions of StARD1, Cyp11a1 and 3-β-HSD mRNAs for KNG cells. ( I ) MDA, GSH and SOD levels were detected for KNG cells. P < 0.05 and P < 0.01 vs. Normal; P < 0.05 and P < 0.01 vs. Control; * P < 0.05 and ** P < 0.01 vs. Model; # P < 0.05 and ## P < 0.01 vs. sh-NC; @ P < 0.05 and @@ P < 0.01 vs. sh-Notch2. Results were presented as mean ± SD

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Knockdown, Cell Counting, CCK-8 Assay, Flow Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: Notch2 acted on the Wnt2/β-catenin pathway in POF cells. ( A ) qPCR analysis for the expressions Wnt2 and β-catenin mRNAs in KNG cells. ( B ) Western blot analysis for the expressions of Notch2, Jag1, Hes2 Wnt2, β-catenin, Axin2 and LEF1 proteins in KNG cells. P < 0.05 and P < 0.01 vs. Control; * P < 0.05 and ** P < 0.01 vs. Model; # P < 0.05 and ## P < 0.01 vs. sh-NC; @ P < 0.05 and @@ P < 0.01 vs. sh-Notch2. Results were presented as mean ± SD

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Western Blot, Control

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: βcatenin knockdown worsened GC functions in POF cells treated with sh-Notch2 and SKL2001 ( A ) CCK-8 assay was used to measure cell viability for KNG cells among groups. ( B ) Flow cytometry was conducted to test cell cycle distribution for KNG cells among groups. ( C ) Flow cytometry was carried out to evaluate cell apoptosis for KNG cells among groups. ( D ) qPCR was performed to detect CDK4, CCND2, P21, Bax and Bcl2 mRNA expressions in KNG cells among groups. ( E ) ELISA was utilized to measure FSH, E2 and AMH levels for KNG cells among groups. ( F ) qPCR was performed to detect StARD1, Cyp11a1 and 3-β-HSD in KNG cells among groups. ( G ) MDA, GSH and SOD levels among groups were detected for KNG cells. P < 0.05 and P < 0.01 vs. Model; # P < 0.05 and ## P < 0.01 vs. sh-Notch2 + SKL2001 + sh-NC. Results were presented as mean ± SD

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Knockdown, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Journal of Ovarian Research

Article Title: Notch2 improves granulosa cell functions in premature ovarian failure by activating the Wnt2/β-catenin pathway

doi: 10.1186/s13048-025-01745-9

Figure Lengend Snippet: βcatenin knockdown inhibited Wnt2/β-catenin pathway in POF cells treated with sh-Notch2 and SKL2001 ( A ) qPCR analysis for the expressions Wnt2 and β-catenin mRNAs in KNG cells. ( B ) Western blot analysis for the expressions of Notch2, Jag1, Hes2 Wnt2, β-catenin, Axin2 and LEF1 proteins in KNG cells. # P < 0.05 and ## P < 0.01 vs. sh-Notch2 + SKL2001 + sh-NC. Results were presented as mean ± SD

Article Snippet: Notch2 , BIOSS , bs-21663R , 1:1000.

Techniques: Knockdown, Western Blot