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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, <t>GSK3β</t> and downstream MYC (Spearman). Representative western blot bands and semi-quantification of protein expression levels <t>of</t> <t>p-GSK3β,</t> 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.
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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.

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: The details of the primary antibodies used were as follows: EFNB2 (1:1,000; cat. no. sc-398735; Santa Cruz Biotechnology, Inc.), Bcl-2 (1:2,000; cat. no. ab182858; Abcam), Bax (1:2,000; cat. no. ab32503; Abcam), CyclinD1 (1:1,000, cat. no. 2978; Cell Signaling Technology, Inc.), CDK4 (1:1,000; cat. no. 12790; Cell Signaling Technology, Inc.), GSK3β (1:5,000; cat. no. 82061-1-RR; Proteintech Group, Inc.), phosphorylated (p-)GSK3β (Ser389; 1:1,000; cat. no. 14850-1-AP; Proteintech Group, Inc.), β-catenin (1:5,000; cat. no. 51067-2-AP; Proteintech Group, Inc.), c-myc (1:5,000; cat. no. 808451-RR; Proteintech Group, Inc.) and β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.).

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