p-gsk3β #14630  (Cell Signaling Technology Inc)

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

Journal: Life

Article Title: The Impact of Decreased GSK3β and S6K1 Expression in TNBC Patients

doi: 10.3390/life15121917

Figure Lengend Snippet: Expression of GSK3β and S6K1 in tumor and normal tissue of TNBC patients (Wilcoxon signed-rank test) in relation to β actin. Median, quartiles maximum, and minimum values are presented.

Article Snippet: Sections were then incubated with the primary antibodies, S6K1 (p70 S6 Kinase Polyclonal Antibody, Invitrogen, Rockford, IL, USA) and GSK3β (GSK3 beta antibody, Biorbyt, Cambridge, UK), according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Life

Article Title: The Impact of Decreased GSK3β and S6K1 Expression in TNBC Patients

doi: 10.3390/life15121917

Figure Lengend Snippet: IHC staining of TNBC tissue: S6K1-cytoplasmic reactivity ( a ); S6K1-positive cytoplasmic and nuclear reactivity ( b ); GSK3β-negative reactivity ( c ); GSK3β-positive focal nuclear reactivity ( d ), The arrow is to see the difference from negative ( c ).

Article Snippet: Sections were then incubated with the primary antibodies, S6K1 (p70 S6 Kinase Polyclonal Antibody, Invitrogen, Rockford, IL, USA) and GSK3β (GSK3 beta antibody, Biorbyt, Cambridge, UK), according to the manufacturer’s instructions.

Techniques: Immunohistochemistry

Journal: Regenerative Therapy

Article Title: The umbilical cord blood exosome MFG-E8 alleviates hypoxic-ischemic encephalopathy brain injury in neonatal rats by restoring autophagy flux and inhibiting ferroptosis through GSK3β/β-catenin signaling

doi: 10.1016/j.reth.2025.06.016

Figure Lengend Snippet: HUBC-Exo-derived MFG-E8 inhibited hyperautophagy and ferroptosis in SH-SY5Y cells that had been subjected to OGD/R through GSK3β/β-catenin signaling. (A) Western blot assessment of p-GSK3β, GSK3β, Active-β-catenin, and β-catenin expression. (B) MFG-E8, p-GSK3β, GSK3β, Active-β-catenin, and β-catenin expression were detected by Western blot. (C) CCK-8 detection of cell viability. (D) Biochemical detection of LDH level in supernatant. (E) Western blot assessment of Beclin1, ATG7, LC3 II/I, and p62 expression. (F) Western blot detection of NCOA4, ACSL4, FTH1, SLC7A11 and GPX4 expression. (G) Biochemical detection of MDA, 4-HNE, and Fe 2+ concentration. (H) Lipid ROS levels assessment of flow cytometry. n = 3. ∗p < 0.05 vs. Control group; #p < 0.05 vs. OGD/R group; &p < 0.05 vs. OGD/R + HUBC-Exo si-NC group; @p < 0.05 vs. OGD/R + HUBC-Exo si-MFG-E8 group.

Article Snippet: GSK3β , #9315 , Rabbit , 1:1000 , 46 KDa , CST , USA.

Techniques: Derivative Assay, Western Blot, Expressing, CCK-8 Assay, Concentration Assay, Flow Cytometry, Control

Journal: Regenerative Therapy

Article Title: The umbilical cord blood exosome MFG-E8 alleviates hypoxic-ischemic encephalopathy brain injury in neonatal rats by restoring autophagy flux and inhibiting ferroptosis through GSK3β/β-catenin signaling

doi: 10.1016/j.reth.2025.06.016

Figure Lengend Snippet: HUBC-Exo-derived MFG-E8 alleviated brain injury in neonatal rats with HIE. (A) Western blot analysis of p-GSK3β, GSK3β, Active-β-catenin, β-catenin expression. (B) Quantitative analysis of cerebral infarction size using TTC staining. (C) The Negative geotaxis reflex test. (D) The Rotarod test, Foot-fault test, and Morris water maze test. (E) Assessment of brain water content. n = 6. ∗p < 0.05 vs. Sham group; #p < 0.05 vs. HIE group; & p < 0.05 vs. HIE + HUBC-Exo si-NC group.

Article Snippet: GSK3β , #9315 , Rabbit , 1:1000 , 46 KDa , CST , USA.

Techniques: Derivative Assay, Western Blot, Expressing, Staining

Journal: Regenerative Therapy

Article Title: Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins

doi: 10.1016/j.reth.2025.05.008

Figure Lengend Snippet: Schema to obtain induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs). (A) A method for iMSC generation via lateral plate mesoderm (LPM). To induce mesodermal lineage, canine iPSCs (ciPSCs) were cultured on Matrigel with 20 % knockout serum replacement (KSR) medium containing transforming growth factor β (TGFβ) signal inhibitor, SB431542 (SB). After passages, the cells were cultured on tissue-culture dishes with fetal bovine serum (FBS)-MSC medium containing basic fibroblast growth factor (bFGF). We described this iMSC induction protocol as the LPM protocol. (B) A method for iMSC generation via neural crest cells (NCCs). To induce NCCs, ciPSCs were cultured with laminine-511 (LN511) and StemFit without solution C (StemFit without C) containing glycogen synthetase kinase 3β (GSK3β) inhibitor, CHIR99031 (CHIR), SB, and bFGF. After neural specification, the cells were maintained in NCC maintenance medium, which was StemFit without C containing SB, bFGF, and epidermal growth factor (EGF) on fibronectin (FN). When NCCs differentiated into iMSCs, they were cultured with StemFit for mesenchymal stem cells and LN511. We described this iMSC induction protocol as NCC + StemFit. (C) iMSC induction method via NCCs using other iMSC culture conditions. NCCs generated as described in (B) were induced to iMSCs by culturing with PRIME-XV MSC expansion XSFM on FN. We described this iMSC induction protocol as NCC + PRIME.

Article Snippet: Briefly, 3–5 days after passage, ciPSCs were cultured in NCC induction medium, which consisted of StemFit without supplement C (StemFit without C) supplemented with 10 ng/mL bFGF, 10 μM SB431542, 1 μM glycogen synthase kinase 3β (GSK3β) inhibitor (CHIR99021; Fujifilm Wako Pure Chemical Corporation) for 10 days at 37 °C and 5 % CO 2 .

Techniques: Derivative Assay, Cell Culture, Knock-Out, Generated