rabbit polyclonal anti β catenin  (Boster Bio)

Journal: Poultry Science

Article Title: Frizzled-4 promotes bone formation in chickens via activation of the canonical Wnt signaling pathway

doi: 10.1016/j.psj.2026.106589

Figure Lengend Snippet: Effects of FZD4 overexpression and knockdown on the expression of key factors in the canonical Wnt signaling pathway. (A-B) Relative expression of signaling pathway key factors C-myc, CyclinD1, β-catenin , and GSK-3β in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (C) Western blot analysis of FZD4, GSK-3β, and β-catenin protein levels at 36 hours post-differentiation induction. (D-E) Band intensity analysis of FZD4, GSK-3β, and β-catenin proteins. n = 3. (F-G) Relative expression of key signaling pathway factors C-myc, CyclinD1, β-catenin , and GSK-3β in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (H) Western blot analysis of FZD4, GSK-3β, and β-catenin protein levels at 36 hours post-differentiation induction. (I-J) Grayscale analysis of FZD4, GSK-3β, and β-catenin protein bands. Band intensity was analyzed using ImageJ software. * P < 0.05, ** P < 0.01. n = 3.

Article Snippet: The primary antibodies used were FZD4 rabbit polyclonal antibody (1:2000, bs-13217R, Bioss), Glycogen synthase kinase 3 beta ( GSK-3β ) rabbit polyclonal antibody (1:2000, 51065-1-AP, Proteintech), β-catenin rabbit polyclonal antibody (1:5000, 51067-2-AP, Proteintech) and GAPDH mouse monoclonal antibody (1:10000, 60004-1-Ig, Proteintech).

Techniques: Over Expression, Knockdown, Expressing, Western Blot, Software

Journal: Poultry Science

Article Title: Frizzled-4 promotes bone formation in chickens via activation of the canonical Wnt signaling pathway

doi: 10.1016/j.psj.2026.106589

Figure Lengend Snippet: Effects of overexpressing FZD4 while inhibiting the canonical Wnt signaling pathway on the osteogenic differentiation of chicken BMSCs. (A-B) Relative expression of osteogenic marker genes Col1A1, Runx2, ALP , and OCN in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (C-D) Relative expression of key signaling pathway factors C-myc, CyclinD1, β-catenin , and GSK-3β in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (E-G) Western blot analysis of β-catenin and GSK-3β protein levels at 36 h post-differentiation induction. Protein band grayscale analysis was performed using ImageJ software. n = 3. (H-I) ALP staining images (Scale bar = 100 μm) and grayscale analysis at 7 days post-induction. n = 4. (J-K) ARS staining (Scale bar = 100 μm) and grayscale analysis at 14 days post-induction. * P < 0.05, ** P < 0.01. n = 4.

Article Snippet: The primary antibodies used were FZD4 rabbit polyclonal antibody (1:2000, bs-13217R, Bioss), Glycogen synthase kinase 3 beta ( GSK-3β ) rabbit polyclonal antibody (1:2000, 51065-1-AP, Proteintech), β-catenin rabbit polyclonal antibody (1:5000, 51067-2-AP, Proteintech) and GAPDH mouse monoclonal antibody (1:10000, 60004-1-Ig, Proteintech).

Techniques: Expressing, Marker, Western Blot, Software, Staining

Journal: Poultry Science

Article Title: Frizzled-4 promotes bone formation in chickens via activation of the canonical Wnt signaling pathway

doi: 10.1016/j.psj.2026.106589

Figure Lengend Snippet: Effects of knockdown FZD4 while inhibiting the canonical Wnt signaling pathway on the osteogenic differentiation of chicken BMSCs. (A-B) Relative expression of osteogenic marker genes Col1A1, Runx2, ALP , and OCN in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (C-D) Relative expression of key signaling pathway factors C-myc, CyclinD1, β-catenin , and GSK-3β in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (E-G) Western blot analysis of β-catenin and GSK-3β protein levels at 36 h post-differentiation induction. Protein band grayscale analysis was performed using ImageJ software. n = 3. (H-I) ALP staining images (Scale bar = 100 μm) and grayscale analysis at 7 days post-induction. n = 4. (J-K) ARS staining (Scale bar = 100 μm) and grayscale analysis at 14 days post-induction. * P < 0.05, ** P < 0.01. n = 4.

Article Snippet: The primary antibodies used were FZD4 rabbit polyclonal antibody (1:2000, bs-13217R, Bioss), Glycogen synthase kinase 3 beta ( GSK-3β ) rabbit polyclonal antibody (1:2000, 51065-1-AP, Proteintech), β-catenin rabbit polyclonal antibody (1:5000, 51067-2-AP, Proteintech) and GAPDH mouse monoclonal antibody (1:10000, 60004-1-Ig, Proteintech).

Techniques: Knockdown, Expressing, Marker, Western Blot, Software, Staining

Journal: Kidney360

Article Title: Saracatinib Delays the Progression of Renal Interstitial Fibrosis by Inhibiting the Wnt/ β -Catenin Pathway

doi: 10.34067/KID.0000000972

Figure Lengend Snippet: Comparison of renal histological characteristics between the normal control group and the CKD group. (A) Results of Masson staining in NHK tissues and kidney tissues of patients with CKD. (B) Immunohistochemistry analysis indicated that Fyn, β-catenin, p-β-catenin, and α-SMA were increased in kidney tissues of patients with CKD compared with NHK tissues. Scale bar: 100 μ m. (**** P < 0.0001, as determined by independent sample t -test). NHK, normal human kidney; α-SMA, α-smooth muscle actin.

Article Snippet: For immunohistochemistry staining, a rabbit monoclonal anti-Fyn antibody (Boster, Wuhan, China, BM4109,1:20 dilution), rabbit polyclonal anti- β -catenin (Boster, Wuhan, China, PA1212, 1:50dilution), rabbit polyclonal anti- p - β -catenin (Tyr142; Abcam, Waltham, MA, ab27798, 1:50 dilution), mouse monoclonal anti-α-smooth muscle actin ( α -SMA) (Cell Signaling Technology, Boston, MA, #69313, 1:100 dilution), biotinylated secondary antibody (Santa Cruz Biotechnology Inc., 1:500), and 3,3′-diaminobenzidine substrate system were used.

Techniques: Comparison, Control, Staining, Immunohistochemistry

Journal: Kidney360

Article Title: Saracatinib Delays the Progression of Renal Interstitial Fibrosis by Inhibiting the Wnt/ β -Catenin Pathway

doi: 10.34067/KID.0000000972

Figure Lengend Snippet: Comparison of renal tissue biological properties between the sham-operated group and the UUO group in rats. (A) IHC analysis revealed that the expression levels of Fyn, β -catenin, p-β -catenin, and α -SMA were elevated in the kidney tissues of the UUO rat model compared with the sham operation group. Scale bar: 100 μ m. (**** P < 0.0001, as determined by independent sample t -test). (B) Western blot analysis of Fyn, β -catenin, p-β -catenin, and α -SMA expression in kidneys from the sham operation group and UUO rat model. Scale bar: 100 μ m. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by independent sample t test). IHC, immunohistochemical.

Article Snippet: For immunohistochemistry staining, a rabbit monoclonal anti-Fyn antibody (Boster, Wuhan, China, BM4109,1:20 dilution), rabbit polyclonal anti- β -catenin (Boster, Wuhan, China, PA1212, 1:50dilution), rabbit polyclonal anti- p - β -catenin (Tyr142; Abcam, Waltham, MA, ab27798, 1:50 dilution), mouse monoclonal anti-α-smooth muscle actin ( α -SMA) (Cell Signaling Technology, Boston, MA, #69313, 1:100 dilution), biotinylated secondary antibody (Santa Cruz Biotechnology Inc., 1:500), and 3,3′-diaminobenzidine substrate system were used.

Techniques: Comparison, Expressing, Western Blot, Immunohistochemical staining

Journal: Kidney360

Article Title: Saracatinib Delays the Progression of Renal Interstitial Fibrosis by Inhibiting the Wnt/ β -Catenin Pathway

doi: 10.34067/KID.0000000972

Figure Lengend Snippet: Comparison of renal tissue biological properties in UUO rats treated with different doses of saracatinib. (A) Results of Masson staining in the UUO rats treated with normal saline, 5-mg saracatinib, and 50-mg saracatinib. (B) IHC staining results for the proteins Fyn, β -catenin, p-β -catenin (Tyr142), and α -SMA in kidney tissue sections from UUO rats treated with NS, 5-mg saracatinib, and 50-mg saracatinib. (C) Western blot analysis of Fyn, β -catenin, p - β -catenin, and α -SMA expression in kidneys from UUO rats treated with NS, 5-mg saracatinib, and 50-mg saracatinib. Scale bar: 100 μ m. (ns implies not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by one-way ANOVA). Tyr142, tyrosine 142.

Article Snippet: For immunohistochemistry staining, a rabbit monoclonal anti-Fyn antibody (Boster, Wuhan, China, BM4109,1:20 dilution), rabbit polyclonal anti- β -catenin (Boster, Wuhan, China, PA1212, 1:50dilution), rabbit polyclonal anti- p - β -catenin (Tyr142; Abcam, Waltham, MA, ab27798, 1:50 dilution), mouse monoclonal anti-α-smooth muscle actin ( α -SMA) (Cell Signaling Technology, Boston, MA, #69313, 1:100 dilution), biotinylated secondary antibody (Santa Cruz Biotechnology Inc., 1:500), and 3,3′-diaminobenzidine substrate system were used.

Techniques: Comparison, Staining, Saline, Immunohistochemistry, Western Blot, Expressing

Journal: Kidney360

Article Title: Saracatinib Delays the Progression of Renal Interstitial Fibrosis by Inhibiting the Wnt/ β -Catenin Pathway

doi: 10.34067/KID.0000000972

Figure Lengend Snippet: Saracatinib inhibits the translocation of β -catenin into nucleus in PTECs. IHC analysis revealed that β -catenin expression in PTECs from sham operation rats, UUO rats, and UUO rats treated with 50-mg saracatinib. Scale bar: 50 μ m. PTEC, primary tubular epithelial cells.

Article Snippet: For immunohistochemistry staining, a rabbit monoclonal anti-Fyn antibody (Boster, Wuhan, China, BM4109,1:20 dilution), rabbit polyclonal anti- β -catenin (Boster, Wuhan, China, PA1212, 1:50dilution), rabbit polyclonal anti- p - β -catenin (Tyr142; Abcam, Waltham, MA, ab27798, 1:50 dilution), mouse monoclonal anti-α-smooth muscle actin ( α -SMA) (Cell Signaling Technology, Boston, MA, #69313, 1:100 dilution), biotinylated secondary antibody (Santa Cruz Biotechnology Inc., 1:500), and 3,3′-diaminobenzidine substrate system were used.

Techniques: Translocation Assay, Expressing

Journal: eLife

Article Title: Wnt induces FZD5/8 endocytosis and degradation and the involvement of RSPO-ZNRF3/RNF43 and DVL

doi: 10.7554/eLife.103996

Figure Lengend Snippet: ( A ) Flow cytometry analysis of FZD levels on the cell surface of WT HEK293A, ZRDKO (ZRDKO C1/2), and HEK293A cells stably expressing V5-FZD5/7 with an anti-pan-FZD monoclonal antibody. FZD levels on the cell surface were greater in ZRDKO cells than in WT cells but much lower than those in V5-FZD5/7-expressing cells. ( B ) Cytosolic β-catenin levels were significantly lower in V5-FZD5/7-expressing cells than in ZRDKO cells. ( C ) ZNRF3/RNF43 double knockout-induced DVL phosphorylation and cytosolic β-catenin accumulation depend on endogenous Wnt proteins. WT or ZRDKO cells were treated with or without IWP-2 and analyzed by immunoblotting. ( D ) Overexpressing V5-FZD5 or V5-FZD7 resulted in Wnt-independent DVL phosphorylation. V5-FZD5/7-expressing cells were treated with or without IWP-2, and the WCLs were analyzed by immunoblotting. ( E ) Expression of ZNRF3 or RNF43, but not RNF130 or RNF150, reversed the increase in cytosolic β-catenin levels in ZRDKO cells. ( F ) Overexpression of RNF43 or RNF150 reduced the levels of mature forms of V5-FZD5 or V5-FZD7. WCLs from HEK293A cells expressing V5-FZD5 or V5-FZD7 alone or together with RNF43-HA or RNF150-HA were analyzed by immunoblotting. ( G ) Overexpression of ZNRF3, RNF43, RNF130, or RNF150 in ZRDKO cells reduced FZD levels on the cell surface, as measured by flow cytometry with an anti-pan-FZD antibody. ( H ) Overexpression of ZNRF3, RNF43, RNF130, or RNF150 in ZRDKO cells reduced FZD5/8 levels on the cell surface. IWP-2 partially reversed the reduction in FZD5/8 levels induced by ZNRF3 or RNF43 but not by RNF130 or RNF150. The cells in ( E ) were treated with or without IWP-2 overnight and analyzed by flow cytometry with an anti-FZD5/8 monoclonal antibody. Figure 5—source data 1. Raw unedited blots for . Figure 5—source data 2. Uncropped and labeled blots for .

Article Snippet: Antibody , Rabbit polyclonal anti-Beta Catenin , Proteintech , Cat# 51067–2-AP; RRID: AB_2086128 , WB(1:10000).

Techniques: Flow Cytometry, Stable Transfection, Expressing, Double Knockout, Phospho-proteomics, Western Blot, Over Expression, Labeling

Journal: eLife

Article Title: Wnt induces FZD5/8 endocytosis and degradation and the involvement of RSPO-ZNRF3/RNF43 and DVL

doi: 10.7554/eLife.103996

Figure Lengend Snippet: ( A ) Flow cytometry analysis of FZD5/8 levels on the cell surface in ZRDKO and ZRDKO-FZD5/8DKO cells with an anti-FZD5/8 antibody. ( B ) Depletion of FZD5/8 diminished the increase in cytosolic β-catenin levels in ZRDKO cells. Cytosolic fractions from the indicated cells were analyzed by immunoblotting. ( C ) Re-expression of V5-FZD5, but not V5-FZD7, restored cytosolic β-catenin levels in ZRDKO-FZD5/8DKO cells. ( D ) Overexpression of V5-FZD, but not V5-FZD7, further elevated cytosolic β-catenin levels in ZRDKO cells. ( E, F ) FZD5, but not FZD7, enhanced the inhibitory effect of RNF43 on Wnt3a signaling. HEK293A cells stably expressing RNF43-HA alone or with V5-FZD5 or V5-FZD7 were treated with increasing doses of Wnt3a for 2 hr, and the cytosolic fractions were analyzed by immunoblotting. ( G ) Flow cytometry analysis of cell surface FZD5/8 levels in WT or FZD5/8 DKO cells. ( H, I ) FZD5/8 double knockout abolished RSPO1-induced cytosolic β-catenin accumulation but had little effect on Wnt3a-induced increases in cytosolic β-catenin levels. The cells were treated with RSPO1 CM for 4 hr ( H ) or increasing doses of Wnt3a for 2 hr ( I ), and the cytosolic fractions were analyzed by immunoblotting. ( J ) WCLs from WT, FZD5/8 DKO and FZD5/8 DKO cells stably expressing V5-FZD5 or V5-FZD8 were analyzed by immunoblotting. ( K ) FZD5/8 double knockout abolished RSPO1-induced cytosolic β-catenin accumulation, which was restored by re-expressing V5-FZD5 or V5-FZD8. Figure 6—source data 1. Raw unedited blots for . Figure 6—source data 2. Uncropped and labeled blots for .

Article Snippet: Antibody , Rabbit polyclonal anti-Beta Catenin , Proteintech , Cat# 51067–2-AP; RRID: AB_2086128 , WB(1:10000).

Techniques: Flow Cytometry, Western Blot, Expressing, Over Expression, Stable Transfection, Double Knockout, Labeling