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: Jag1 , BIOSS , bs-1448R , 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 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: Jag1 , BIOSS , bs-1448R , 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 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: Jag1 , BIOSS , bs-1448R , 1:1000.

Techniques: Knockdown, Western Blot

Journal: bioRxiv

Article Title: The Notch1 intracellular domain orchestrates mechanotransduction of fluid shear stress

doi: 10.1101/2025.07.13.663563

Figure Lengend Snippet: (A) Western blot of hdBEC lysates cultured under static or flow (∼20 dynes/cm ) conditions, with pre-treatment with 0.5 μg/mL of rhDll4 or vehicle control. (B) Quantification of normalized Western blot intensity of Notch1 V1754. (C) Western blot of hdBEC lysates from scramble (SCR), DLL4 KO , and JAG1 KO hdBECs under static and flow conditions. (D) Quantification of normalized Western blot intensity of Notch1 V1754. (E) Western blot of hdBEC and hdLEC lysates under static and flow conditions. (F) Quantification of normalized Western blot intensity of Notch1 V1754. (G) Fluorescence micrographs of hdBECs and hdLECs under static and flow conditions immunostained for Notch1 ICD (heatmap) and VE-cadherin (white). Scale bar, 25 μm. (H) Quantification of the relative Notch1 polarization in hdBECs versus hdLECs under flow. n ≥ 10 fields of view from three independent experiments. (I) Timelapse of Notch1-GFP in hdBEC cells under flow. Time scale (min:sec). Scale bar, 10 μm. (J) Fluorescence micrographs of SCR and DLL4 KO cells under flow conditions immunostained for Notch1 (black) and DNA (blue). Scale bar, 25 μm. (K) Quantification of the relative degree of Notch1 polarization in SCR versus DLL4 KO cells under flow. n ≥ 10 fields of view from three independent experiments. (K) Quantification of the relative degree of ligand polarization in Dll4-GFP or Jag1-mEmerald cells under flow. n ≥ 10 fields of view from three independent experiments. Western blots are representative of three independent experiments. For all plots, mean ± SD; one-way Anova with Tukey’s post-hoc test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns denotes non-significant.

Article Snippet: Antibodies against Notch1 V1754 (V1744 in mice, D3B8, 1:500 WB), Notch1 ICD (D1E11, 1:200 IF, 1:1000 WB), GAPDH (14C10, 1:5000 WB), Dll4 (D7N3H, 1:1000 WB), Jag1 (D4Y1R, 1:1000 WB), GFP (D5.1, 1:5000 WB), Annexin A2 (D11G2, 1:1000 WB, 1:200 IF), Presenilin-1 (E3L9X, 1:1000 WB), and Flotillin-2 (C42A3, 1:1000 WB) were from Cell Signaling Technologies.

Techniques: Western Blot, Cell Culture, Control, Fluorescence