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: Isolation, Culture, and In Vitro Osteogenic Differentiation of Chicken BMSCs. (A) Morphological characteristics of BMSCs, (1) - (5) represent features at 0, 1, 5, 7, and 9 days post-culture, respectively. (Scale bar = 50 μm) (B) Immunofluorescence; CD29: Also known as beta-1 integrin, a positive surface marker for mesenchymal stem cells. CD45: Also known as Leukocyte Common Antigen (LCA), a hematopoietic cell marker. (C) ALP staining and ARS staining images after induced differentiation (Scale bar = 100 μm). (D-E) Relative expression of osteogenic differentiation marker gene ALP and target gene FZD4 at 0 and 3 days post-induced differentiation. n = 3. (F) Western blot analysis of FZD4 protein levels at 36 hours post-induced differentiation. (G) Gray-scale analysis of FZD4 protein bands. ImageJ software was used for band analysis. * 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: Isolation, In Vitro, Immunofluorescence, Marker, Staining, 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 FZD4 overexpression and knockdown on the osteogenic differentiation of chicken BMSCs. (A) FZD4 expression in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 3. (B-C) Relative expression of osteogenic marker genes Col1A1, Runx2, ALP , and OCN in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (D-E) ALP staining and grayscale analysis at 7 days post-induction. n = 4. (F-G) ARS staining and grayscale analysis at 14 days post-induction. Grayscale analysis was performed using ImageJ software. n = 4. (H) FZD4 expression in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 3. (I-J) Relative expression of osteogenic marker genes Col1A1, Runx2, ALP , and OCN in chicken BMSCs at 0 and 7 days post-osteogenic differentiation. n = 6. (K-L) ALP staining and grayscale analysis at 7 days post-induction (Scale bar = 100 μm). n = 4. (M-N) ARS staining and grayscale analysis at 14 days post-induction (Scale bar = 100 μm). 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: Over Expression, Knockdown, Expressing, Marker, Staining, 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 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