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

Journal: Biochemistry and Biophysics Reports

Article Title: Comparative analysis of activation of macrophages/microglia in diabetic retinopathy and Familial Exudative Vitreoretinopathy

doi: 10.1016/j.bbrep.2025.102396

Figure Lengend Snippet: FZD4 knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.

Article Snippet: Knockout of FZD4 in these mice were validated by Western blot for FZD4 in the retinal tissue, using monoclonal antibody against mouse FZD4 (MAB195–050, R&D Systems).

Techniques: Knock-Out, Western Blot

Journal: bioRxiv

Article Title: Cell-autonomous Wnt activity promotes transient re-programming and cell cycle re-entry of coronary artery endothelial cells

doi: 10.64898/2026.02.23.707374

Figure Lengend Snippet: ( A ) Violin plot showing levels of mRNA expression of Frizzled (Fzd) receptors in neonatal hearts. ( B ) Cryosection from Cx40CreER; Rosa26 TdTomato heart showing a cross-section of a coronary artery, immunostained for Fzd4 (green). TdTomato + lineage traced arterial endothelial cells shown in red, nuclei labelled with DAPI in blue. ( C ) Experimental design to assess the role of arterial Fzd4 in artery reassembly. ( D - F ) Confocal images of neonatal Cx40CreER; Rosa26 TdTomato hearts labelled and traced for pre-existing artery endothelial cells in black. D shows whole heart. ( E , F ) Images of watershed from control hearts with ( E ) moderate (inset from D ) and ( F ) severe MI. ( G ) Quantification of TdTomato + collaterals per heart. p<0.0001. ( H - J ) Confocal images of neonatal Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato hearts, labelled and traced for pre-existing artery endothelial cells (shown in black). H shows whole heart. ( I , J ) Images of watershed from control hearts with ( I ) moderate (inset from H ) and ( J ) severe MI. Arrowheads and brackets point to single and clusters of saECs, respectively. All 5 knockout hearts undergoing moderate MI demonstrate a phenotype as shown in I . 4 out of 6 knockout hearts undergoing severe MI demonstrate a phenotype as shown in J. ( K ) Confocal image of Fzd4 depleted single artery cells, post-MI show non-EC morphology. ( L , M ) Confocal images of watershed regions from ( L ) control and ( M ) arterial knockouts for Fzd4, showing EdU + proliferating cells in green and TdTomato + lineage traced artery endothelial cells in red. Arrowheads point to EdU + TdTomato + proliferating single artery cells, post-MI. ( N ) Quantification of EdU + proliferating single artery cells post-MI. p=0.0186. Scale bars: B , 50µm; D , H , 500µm; E , F , I , J , 200 µm; K - M , 50 µm. trans, transient; cyc, cycling; P, postnatal day; EC, endothelial cell; Tam, Tamoxifen; MI, myocardial infarction; lig, ligation; 1°, primary; LCA, left coronary artery; CM, cardiomyocyte; saEC, single artery endothelial cell

Article Snippet: Following mouse lines, Cx40CreER , ApjCreER , Cdk1 fl/fl (The Jackson Laboratory, 129S(B6N)-Cdk1 tm1Eddy /J, stock number 028028), Fzd4 fl/fl (The Jackson Laboratory, B6;129-Fzd4 tm2.1Nat /J, stock number 011078), Wls fl/fl (The Jackson Laboratory, Wls tm1.1Whsu /J, stock number 027484), Rosa26 TdTomato Cre reporter line (The Jackson Laboratory, B6.Cg-Gt[ROSA]26Sortm9[CAG-TdTomato]Hze/J, stock number 007909), Cx40 eGFP/+ and Axin2-d2EGFP were used in the experiments.

Techniques: Expressing, Control, Knock-Out, Ligation

Journal: bioRxiv

Article Title: Cell-autonomous Wnt activity promotes transient re-programming and cell cycle re-entry of coronary artery endothelial cells

doi: 10.64898/2026.02.23.707374

Figure Lengend Snippet: ( A ) Secondary alone control for immunostaining shown in . ( B ) Schematic showing positions of coronary occlusion through ligation of main LCA or primary branches of LCA to generate severe or moderate myocardial infarctions, respectively. ( C , D ) Confocal images of watershed regions from Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato hearts with severe MI. Arrowheads point to TdTomato + lineage traced endothelial cells with non-EC morphologies. Brackets point to cluster of TdTomato + endothelial cells. ( E - G ) Confocal images of TdTomato + endothelial cells traced from pre-existing arteries showing non-EC morphology post-severe MI, in Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato hearts. * point to TdTomato-lineage traced cells with cardiomyocyte like features. Scale bars: A , E , F , G , 50µm; C , D , 200µm. 1°, primary; 2°, secondary; LCA, left coronary artery; RCA, right coronary artery; EC, endothelial cell; MI, myocardial infarction; Ab, Antibody.

Article Snippet: Following mouse lines, Cx40CreER , ApjCreER , Cdk1 fl/fl (The Jackson Laboratory, 129S(B6N)-Cdk1 tm1Eddy /J, stock number 028028), Fzd4 fl/fl (The Jackson Laboratory, B6;129-Fzd4 tm2.1Nat /J, stock number 011078), Wls fl/fl (The Jackson Laboratory, Wls tm1.1Whsu /J, stock number 027484), Rosa26 TdTomato Cre reporter line (The Jackson Laboratory, B6.Cg-Gt[ROSA]26Sortm9[CAG-TdTomato]Hze/J, stock number 007909), Cx40 eGFP/+ and Axin2-d2EGFP were used in the experiments.

Techniques: Control, Immunostaining, Ligation

Journal: bioRxiv

Article Title: Cell-autonomous Wnt activity promotes transient re-programming and cell cycle re-entry of coronary artery endothelial cells

doi: 10.64898/2026.02.23.707374

Figure Lengend Snippet: ( A ) Experimental design to assess expression of VegfR2 and endomucin in single artery endothelial cells (saECs), post-MI. ( B , C ) Quantification of ( B ) VegfR2 and ( C ) Endomucin, in saECs, post-MI. p<0.0001. Each data point is a TdTomato + single artery cell. N=3 and N=5 hearts were quantified for VegfR2 and Endomucin expression, respectively. ( D ) Confocal images of saECs in Cx40CreER; Rosa26 TdTomato ; Cx40 eGFP/+ hearts, post-MI, showing TdTomato + saECs in red and expression of eGFP as a reporter for Cx40 expression in green. Arrowheads point to TdTomato + saECs with low levels of eGFP signal, as compared to the artery tip. ( E , H ) Confocal images of post-MI watersheds, showing TdTomato + saECs in red and immunostaining for VegfR2 in green in ( E ) control ( Cx40CreER; Rosa26 TdTomato ) and ( F ) knockout ( Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato ) MI hearts. F , G are insets from E and show TdTomato + saECs express VegfR2 (orange arrowheads). I , J are insets from H and show reduced expression of VegfR2 by TdTomato + saECs (magenta arrowheads). ( K , N ) Confocal images of post-MI watersheds, showing TdTomato + saECs in red and immunostaining for Endomucin in green in ( K ) control ( Cx40CreER; Rosa26 TdTomato ) and ( N ) knockout ( Fzd4 L/L ; Cx40CreER; Rosa26 TdTomato ) MI hearts. L , M are insets from K and show TdTomato + saECs express Endomucin (orange arrowheads). O , P are insets from N and show reduced expression of Endomucin by TdTomato + saECs (magenta arrowheads). Scalebars: 50µm. saECs, single artery endothelial cells; P, postnatal day; KO, knockout; cap, capillary; MI, myocardial infarction.

Article Snippet: Following mouse lines, Cx40CreER , ApjCreER , Cdk1 fl/fl (The Jackson Laboratory, 129S(B6N)-Cdk1 tm1Eddy /J, stock number 028028), Fzd4 fl/fl (The Jackson Laboratory, B6;129-Fzd4 tm2.1Nat /J, stock number 011078), Wls fl/fl (The Jackson Laboratory, Wls tm1.1Whsu /J, stock number 027484), Rosa26 TdTomato Cre reporter line (The Jackson Laboratory, B6.Cg-Gt[ROSA]26Sortm9[CAG-TdTomato]Hze/J, stock number 007909), Cx40 eGFP/+ and Axin2-d2EGFP were used in the experiments.

Techniques: Expressing, Immunostaining, Control, Knock-Out

Journal: The Journal of Biological Chemistry

Article Title: Sphingosine-1-phosphate receptor 2 inhibition ameliorates familial exudative vitreoretinopathy models

doi: 10.1016/j.jbc.2025.111107

Figure Lengend Snippet: FZD4 inhibition prevents primary human retinal microvascular endothelial cell (hRMEC) tubularization. A , Microscopy of tubularization in primary hRMECs treated with FZD4 shRNA. B , computational analysis of vascularization in both FZD4 shRNA treated and control. Eight different parameters were examined and measured using FIJI ( , ). C , microscopy of tubularization in hRMECs due to expression of the patient derived dominant-negative FZD4 Met493-Trp49del allele. D , computational analysis of vascularization in both overexpression of the FZD4 variant and the control. There is significant reduction in all eight examined parameters when the FZD4 variant is overexpressed. Ordinary one-way ANOVAs with multiple comparisons were performed on the data. ∗ p < 0.05, ∗∗ p < 0.01; ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Size bar is 500 μM.

Article Snippet: Human shRNA Plasmid Kit (Origene, catalog number TL312883) was used for shRNA knockdown of FZD4 expression.

Techniques: Inhibition, Microscopy, shRNA, Control, Expressing, Derivative Assay, Dominant Negative Mutation, Over Expression, Variant Assay

Journal: The Journal of Biological Chemistry

Article Title: Sphingosine-1-phosphate receptor 2 inhibition ameliorates familial exudative vitreoretinopathy models

doi: 10.1016/j.jbc.2025.111107

Figure Lengend Snippet: The S1PR inhibitor JTE-013 rescues FZD4 mediated primary human retinal microvascular endothelial cell (hRMEC) tubularization defects. A , microscopy of vascular formation in the primary hRMECs treated with FZD4 shRNA and the control (shRNA scramble) with and without JTE-013. Cells were seeded onto a basement membrane and incubated for 6 h at 37 °C prior to imaging. JTE-013 was added at the time of seeding. B , parameters measured by the angiogenesis plugin on FIJI and their statistical analysis. Ordinary one-way ANOVAs with multiple comparisons were performed on the data. ∗ p < 0.05, ∗∗ p < 0.01; ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Size bar is 500 μM.

Article Snippet: Human shRNA Plasmid Kit (Origene, catalog number TL312883) was used for shRNA knockdown of FZD4 expression.

Techniques: Microscopy, shRNA, Control, Membrane, Incubation, Imaging

Journal: Scientific Reports

Article Title: A novel variant p.Y250C of FZD4 influences Norrine/β-catenin signaling pathway that associates with familial exudative vitreoretinopathy (FEVR)

doi: 10.1038/s41598-025-34442-0

Figure Lengend Snippet: Construction and verification of mutant FZD4 plasmids. ( A ) A schematic illustration of human FZD4 protein is presented, with numbers donating the amino acid positions within the specified domains of FZD4. The mutation located in the first intracellular loop, as the arrow indicated. SPs: Signal peptides. CRD: Cysteine-rich domain, which serves as the ligand-binding domain. TM: Single pass of the transmembrane domain. ( B ) The electrophoresis results reveal the FZD4 amplified plasmid and target DNA fragment after enzyme digestion. Lanes 1–3 lanes exhibit all digestion products from the FZD4 plasmid, with DNA fragment sizes of 8000 bp (the black arrow indicated) and 179 bp (the white arrow indicated) after digestion, respectively; lane 4 displays the electrophoresis outcome of 1 µl target DNA fragment, confirming that the substitution DNA fragment obtained post-enzyme digestion was 179 bp (the white arrow indicated) in size. ( C) The results of electrophoresis for 1 µl of each expression vector DNA fragment (the black arrow indicated) in lane 1 and the substitution DNA fragment (the white arrow indicated) in lane 2 before connection serves as a reference for the preparation of the ligation system. ( D and E ) Partial PCR identification results of the bacterial solutions. Three positive clones were identified from the PCR electrophoresis results: No. 9, No. 104 and No. 107, as indicated by arrows. Sequencing results in ( F and G ) confirm that the positive clone plasmids No. 9 and No. 104 contain reverse substitution sequences carrying mutation sites. ( H) The positive clone plasmid No. 107 possesses a forward substitution sequence harboring the mutation. ( M ) DNA molecular size marker DL5000. The arrows signify the mutation sites. Full-length blots are included in the Supplementary Fig. .

Article Snippet: The wild-type FZD4 plasmid (GFP-tagged) (FZD4-WT) was sourced from OriGene (USA) (cat. : RG217286).

Techniques: Mutagenesis, Ligand Binding Assay, Electrophoresis, Amplification, Plasmid Preparation, Expressing, Ligation, Clone Assay, Sequencing, Marker

Journal: Scientific Reports

Article Title: A novel variant p.Y250C of FZD4 influences Norrine/β-catenin signaling pathway that associates with familial exudative vitreoretinopathy (FEVR)

doi: 10.1038/s41598-025-34442-0

Figure Lengend Snippet: Detection of wild-type and mutant FZD4 on downstream signal proteins by luciferase reporter gene assay. FZD4-wild-type (FZD4-WT) and FZD4-mutant (FZD4-MU) represent the relative luciferase activity ascertained from the transfection of wild-type FZD4 plasmid and mutant FZD4 plasmid into Hela cells, respectively. Data are representative of three independent experiments. The experimental data are presented as the mean ± standard deviation (mean ± SD). Differences between groups were assessed using the Student’s t -test, and * P < 0.05.

Article Snippet: The wild-type FZD4 plasmid (GFP-tagged) (FZD4-WT) was sourced from OriGene (USA) (cat. : RG217286).

Techniques: Mutagenesis, Luciferase, Reporter Gene Assay, Activity Assay, Transfection, Plasmid Preparation, Standard Deviation

Journal: Scientific Reports

Article Title: A novel variant p.Y250C of FZD4 influences Norrine/β-catenin signaling pathway that associates with familial exudative vitreoretinopathy (FEVR)

doi: 10.1038/s41598-025-34442-0

Figure Lengend Snippet: Immunostaining for FZD4 and Calnexin. The immunostaining for wild-type or mutant FZD4 and Calnexin in Hela cells. FZD4 protein shows in the green image on the left side of the panel; Calnexin displays with red image; nucleus shows in the blue image; the merge of them shows on the right side. Scale bar: 10 μm. Magnification: × 50.

Article Snippet: The wild-type FZD4 plasmid (GFP-tagged) (FZD4-WT) was sourced from OriGene (USA) (cat. : RG217286).

Techniques: Immunostaining, Mutagenesis

Journal: Scientific Reports

Article Title: A novel variant p.Y250C of FZD4 influences Norrine/β-catenin signaling pathway that associates with familial exudative vitreoretinopathy (FEVR)

doi: 10.1038/s41598-025-34442-0

Figure Lengend Snippet: Subcellular localizations and nuclear entry ratios of wild-type and mutant FZD4 proteins. ( A – C ) Subcellular localization of wild-type FZD4 protein express throughout the cytoplasm in HeLa cells, without highlights. ( D – F ) Subcellular localization of wild-type FZD4 protein aggregate on cell membrane and cytoplasm of HeLa cells, with highlights. ( G – I ) Subcellular localization of mutant FZD4 protein express throughout the cytoplasm in HeLa cells, without highlights. ( J – L ) Subcellular localization of mutant FZD4 protein aggregate on nucleic membrane and nucleus of HeLa cells, with highlights. The highlights represent the aggregation of GFP-labeled FZD4 protein. FZD4 protein shows in the green image on the left side of the panel; nucleus shows in the blue image in the middle; the merge of them shows on the right side. M: The ratio of wild-type FZD4 and mutant FZD4 proteins entering the nucleus. FZD4-WT represents the proportion of wild-type FZD4 protein entering the nucleus, while FZD4-MUT denotes the proportion of mutant FZD4 protein entering the nucleus. * P < 0.05. Scale bar: 5 μm. Magnification: × 50.

Article Snippet: The wild-type FZD4 plasmid (GFP-tagged) (FZD4-WT) was sourced from OriGene (USA) (cat. : RG217286).

Techniques: Mutagenesis, Membrane, Labeling