Journal: International Journal of Oncology

Article Title: Ephrin-B2 promotes gastric cancer growth by inhibiting apoptosis and regulating the cell cycle via the Wnt/β-catenin signaling pathway

doi: 10.3892/ijo.2025.5821

Figure Lengend Snippet: Wnt/β-catenin signaling pathway is responsible for EFNB2-mediated biological effects in gastric cancer cells. (A) Gene Expression Profiling Interactive Analysis and (B) Tumor Immune Estimation Resource databases were employed to explore the transcriptional correlation between EFNB2 and CTNNB1, GSK3β and downstream MYC (Spearman). Representative western blot bands and semi-quantification of protein expression levels of p-GSK3β, GSK3β, β-catenin and c-myc detected under the condition of EFNB2 (C) knockdown and (D) overexpression (one-way ANOVA for AGS cells; unpaired Student's test for HGC-27 cells). Representative western blot bands and semi-quantification of protein expression levels in the presence of (E) an agonist (CHIR99021) and (F) an inhibitor (DIF-3) of the Wnt/β-catenin signaling pathway in the EFNB2 knockdown or overexpression groups, respectively. Protein levels of p-GSK3β, GSK3β, β-catenin and c-myc were detected by western blotting (one-way ANOVA). Data are presented as the mean ± SD. * P<0.05, ** P<0.01, *** P<0.001 vs. sh-NC or vector group. ns, not significant; NC, negative control; DIF-3, differentiation-inducing factor-3; p-, phosphorylated; OE, overexpression vector; sh, short hairpin RNA; CTNNB1, catenin β1; TPM, transcript per million; EFNB2, ephrin-B2.

Article Snippet: For the rescue experiments, cells in designated groups were treated at 37°C for 24 h with either CHIR99021 (5 μ M; MedChemExpress) to activate the Wnt/β-catenin signaling cascade or DIF-3 (30 μ M; MedChemExpress) to inhibit the Wnt/β-catenin signaling cascade.

Techniques: Gene Expression, Western Blot, Expressing, Knockdown, Over Expression, Plasmid Preparation, Negative Control, shRNA

Journal: International Journal of Oncology

Article Title: Ephrin-B2 promotes gastric cancer growth by inhibiting apoptosis and regulating the cell cycle via the Wnt/β-catenin signaling pathway

doi: 10.3892/ijo.2025.5821

Figure Lengend Snippet: EFNB2 promotes the tumor growth of gastric cancer via the Wnt/β-catenin signaling pathway in vivo . (A) Tumors from the immunodeficient nude mice in the four experimental groups were excised and imaged. (B) Subcutaneous tumor growth (mm 3 ) was measured every 3-4 days following various treatments (one-way ANOVA). (C) Excised tumor masses (g) were weighed and compared across groups (one-way ANOVA). (D) Histopathological evaluation of xenograft tumors included H&E staining to examine tissue morphology, IHC analysis of EFNB2, β-catenin and Ki67 expression, and TUNEL staining for apoptotic cell evaluation. All images were captured at a magnification of ×200. (E) Quantification of IHC scores for EFNB2 and β-catenin, and the percentage of positive cells for Ki67 and TUNEL staining (Kruskal-Wallis H test for EFNB2 and β-catenin; one-way ANOVA for Ki67 and TUNEL staining). Data are presented as the mean ± SD. * P<0.05, ** P<0.01, *** P<0.001. ns, not significant; EFNB2, ephrin-B2; DIF-3, differentiation-inducing factor-3; OE, overexpression vector; IHC, immunohistochemical.

Article Snippet: For the rescue experiments, cells in designated groups were treated at 37°C for 24 h with either CHIR99021 (5 μ M; MedChemExpress) to activate the Wnt/β-catenin signaling cascade or DIF-3 (30 μ M; MedChemExpress) to inhibit the Wnt/β-catenin signaling cascade.

Techniques: In Vivo, Staining, Expressing, TUNEL Assay, Over Expression, Plasmid Preparation, Immunohistochemical staining

Journal: EMBO Reports

Article Title: Cyst-independent oocyte phagocytosis builds the female reproductive reserve in mice

doi: 10.1038/s44319-025-00663-7

Figure Lengend Snippet: ( A ) The analysis of oocyte phagocytosis and development in cultured ovaries treated from c-17.5 to c-PD4 with autophagy inhibitors (3MA, MRT68921, BafA1, and CQ). Autophagy inhibition consistently reduced sacrifice and phagocytosis at c-PD1 and increased the survival of oocytes by c-PD4. Scale bar: 20 μm. ( B ) Statistical evaluations showed that suppression of autophagy significantly diminishes oocyte sacrifice and phagocytosis at c-PD1, demonstrating a sharp decline in the ratio of surviving oocytes with FLs and a decrease in the formation of ODs from sacrificed oocytes. The analysis included four ovaries per group, evaluating more than 70 oocytes per ovary. FL: control vs. 3-MA: p value = 5.9E-09; control vs. MRT: p value = 6.7E-08; control vs. BafA1: p value = 1.9E-08; control vs. CQ: p value = 1.1E-06; 3-MA vs. MRT: p value = 0.21; 3-MA vs. BafA1: p value = 0.79; 3-MA vs. CQ: p value = 0.003; MRT vs. BafA1: p value = 0.80; MRT vs. CQ: p value = 0.22; BafA1 vs. CQ: p value = 0.05. OD: control vs. 3-MA: p value = 2.3E-10; control vs. MRT: p value = 4.1E-10; control vs. BafA1: p value = 7.2E-09; control vs. CQ: p value = 7.8E-08; 3-MA vs. MRT: p value = 0.96; 3-MA vs. BafA1: p value = 0.54; 3-MA vs. CQ: p value = 0.66; MRT vs. BafA1: p value = 0.23; MRT vs. CQ: p value = 0.94; BafA1 vs. CQ: p value = 0.07. ( C , D ) Immunofluorescent staining of the ovarian morphology ( C ) and oocyte number counting ( D ) at c-PD4 showed an increased number of surviving oocytes in ovaries treated with autophagy inhibitors. Red - DDX4; Green - FOXL2; Blue - Hoechst. Scale bar: 100 μm. Data were from at least three ovaries per group. Data were presented as the mean ± SD. control vs. 3-MA: p value = 7.25E-05; control vs. MRT: p value = 0.034; control vs. BafA1: p value = 0.011; control vs. CQ: p value = 0.0054; 3-MA vs. MRT: p value = 0.012; 3-MA vs. BafA1: p value = 0.035; 3-MA vs. CQ: p value = 0.155; MRT vs. BafA1: p value = 0.96; MRT vs. CQ: p value = 0.71; BafA1 vs. CQ: p value = 0.95. Statistical significance was determined by ANOVA tests. P (a, b) < 0.05, P (a, c) < 0.05, P (b, c) < 0.05.

Article Snippet: In the autophagy inhibitors or Formin inhibitor treatment experiments, after 4 days of adherent culture, the Oct4-CreER T2 ;mTmG or Oct4-CreER T2 ;Rainbow ovaries at c-17.5 dpc were treated with or without autophagy signaling cascade inhibitors (5 mM 3-MA, M9281, Sigma-Aldrich; 400 nM MRT68921, S7949, Selleckchem; 0.2 nM Bafilomycin A1, HY-100558, MedChemExpress; 4 μM Chloroquine, HY-17589A, MedChemExpress) or Formin inhibitor (20 μM SMIFH2, HY-16931, MedChemExpress) for 3 or 7 days, which media was changed every other day.

Techniques: Cell Culture, Inhibition, Control, Staining