Journal: Stem cells and development

Article Title: Notch-Induced Expression of FZD7 Requires Noncanonical NOTCH3 Signaling in Human Breast Epithelial Cells.

doi: 10.1089/scd.2015.0315

Figure Lengend Snippet: FIG. 1. NOTCH3 regulates expression of Frizzled7 (FZD7) (A) The human breast epithelial cell strain (HMECs) were transduced with different lenti-shNOTCH1- 4 virus and transcript expression of FZD7, lipoprotein receptor-related protein 5 (LRP5) and delta-like 3 (DLL3) were examined using qPCR and normalized with respect to GAPDH and then to the scrambled control-infected cells. (B) 184-hTert cells were infected separately with different lenti-ShNOTCH1-4 or a scrambled control expressing virus and FZD7 transcript expression was determined as in (A). As shown, the transcript expression of FZD7 is only de- creased when NOTCH3 expression is decreased. (n = 3, *P = £0.05, **P = £0.005, ***P = £0.0005).

Article Snippet: For this purpose, specific antibodies raised against FZD7 (1:100 dilution; R&D Systems clone# 151143), NOTCH1 (Cat No. A301-894A; Bethyl Laboratories), NOTCH2 (Cat No. A301-083A; Bethyl Laboratories), and NOTCH3 (Cat No. 2889; Cell Signaling) were utilized.

Techniques: Expressing, Transduction, Virus, Control, Infection

Journal: Stem cells and development

Article Title: Notch-Induced Expression of FZD7 Requires Noncanonical NOTCH3 Signaling in Human Breast Epithelial Cells.

doi: 10.1089/scd.2015.0315

Figure Lengend Snippet: FIG. 2. FZD7 is a unique target of NOTCH3 184-hTert cells and was infected with lentivirus expressing the constitutively active form of each Notch receptor (NR1-4) or empty vector (EV) control and the transcript expression of FZD7 was examined via qPCR (A). These data are quantified relative to the GAPDH expression and is presented relative to the EV control transcript expression. As shown, only increased signaling through NOTCH3 increased FZD7 expression. (B) Flow cytometer was used to examine FZD7 protein expression levels in 184-hTert cells separately transduced to express different Notch receptors (NR1-4) or transduced with different lenti-shNOTCH (shNR1-4) to knockdown expression of each receptor sepa- rately. Some cells were transduced with a lenti-shScrambled-expressing virus (scr) as controls. Representative histograms are shown and average (N = 3) median fluorescent intensities are depicted as bar graphs (shNOTCH-infected cells in C and lenti NR-infected cells in D). As shown, FZD7 protein expression is only altered when signaling through NOTCH3 is modulated. The error bars show the standard deviation ***P = £0.0005). Color images available online at www.liebertpub.com/scd

Article Snippet: For this purpose, specific antibodies raised against FZD7 (1:100 dilution; R&D Systems clone# 151143), NOTCH1 (Cat No. A301-894A; Bethyl Laboratories), NOTCH2 (Cat No. A301-083A; Bethyl Laboratories), and NOTCH3 (Cat No. 2889; Cell Signaling) were utilized.

Techniques: Infection, Expressing, Plasmid Preparation, Control, Flow Cytometry, Transduction, Knockdown, Virus, Standard Deviation

Journal: Stem cells and development

Article Title: Notch-Induced Expression of FZD7 Requires Noncanonical NOTCH3 Signaling in Human Breast Epithelial Cells.

doi: 10.1089/scd.2015.0315

Figure Lengend Snippet: FIG. 3. Luminal progenitors show high expression of FZD7 and NOTCH3 (A) To examine the expression level of FZD7 in the undifferentiated subpopulations of human breast epithelial cells, luminal and the stem/bipotential progenitor-enrich cell populations were obtained from breast reduction samples based on their expression of Epithelial Cell Adhesion (EpCAM) molecule and a-6 Integrin (CD49f). (B,C) Flow cytometry was used to examine FZD7 protein expression in the luminal progenitor (LP)-enriched population (EpCAMbrightCD49flow) and the stem/bipotential progenitor (BP)-enriched subpopulation (EpCAMlowCD49fbright). As shown, the LP subpopulation was more enriched for FZD7-expressing cells and that they also expressed FZD7 at a higher level compared with BP (20.56 – 3.3, 12.23 – 1.3 respectively). (D) RNA was extracted from LP and the BP subpopulations isolated from three different breast reduction samples and the FZD7 and NOTCH3 transcript levels were examined using qPCR. As shown, LP cells express FZD7 and NOTCH3 transcripts at a higher level compared to the BP cells. The transcript levels are normalized to GAPDH transcript levels and then normalized to BP transcript expression levels (*P = £0.05, ***P = £0.0005). (E) Breast reduction samples (N = 3) where dissociated into single cells and infected either with lentivirus expressing empty vector (EV) or NR3 expressing vector. After 72 h, cells expressing the green fluorescent protein (GFP+) cells were examined for their expression of FZD7 via flow cytometry. One representative histogram is shown depicting FZD7 expression in the lenti-EV or -NR3-transduced cells and the average median fluorescence intensity of three samples is shown with standard deviations (**P = £0.005). Color images available online at www.liebertpub.com/scd

Article Snippet: For this purpose, specific antibodies raised against FZD7 (1:100 dilution; R&D Systems clone# 151143), NOTCH1 (Cat No. A301-894A; Bethyl Laboratories), NOTCH2 (Cat No. A301-083A; Bethyl Laboratories), and NOTCH3 (Cat No. 2889; Cell Signaling) were utilized.

Techniques: Expressing, Flow Cytometry, Isolation, Infection, Plasmid Preparation, Cytometry

Journal: Stem cells and development

Article Title: Notch-Induced Expression of FZD7 Requires Noncanonical NOTCH3 Signaling in Human Breast Epithelial Cells.

doi: 10.1089/scd.2015.0315

Figure Lengend Snippet: FIG. 4. NOTCH3 regulates FZD7 expression in a CBF1-Suppressor of Hairless-Lag-1-independent manner (A) 184- hTERT cells were transduced with a NOTCH3-FLAG (NR3-FLAG) expressing constructs and chromatin immunopre- cipitation (ChIP) was performed using anti-FLAG antibody. FLAG only expressing cells were used as controls and the data obtained from these cells are depicted in the Supplementary Figure S5A. The NR3 binding and enrichment on FZD7 promoter on two putative CSL binding sites was assessed via qPCR compared IgG isotype control (set to1). Using primers flanking the CSL binding sites, no significant enrichment of NR3 on FZD7 promoter could be detected. As a positive control the enrichment for binding of NOTCH1 (NR1) and NR3 on the HES1 promoter was also examined. (B,C) 184-hTERT cells overexpressing NR3 or empty vector (EV) control were additionally transduced with lenti-shCSL or lenti-Scrambled control (scr) and transcript expression of CSL and HES1 (B), NOTCH3 and FZD7 (C) was examined via qPCR. These data are quantified relative to the GAPDH expressions and then to the EV-Scr (set to 1). The lenti-shCSL-transduced 184-hTert cells showed >90% reduction in CSL transcript expression. (D) Flow cytometry was used to examine FZD7 protein expression in the EV-CSL-knockdown (EV-shCSL), EV-Scr control (EV-Scr), NR3-overexpressing-CSL-knockdown (NR3-shCSL), and the NR3-overexpressing-Scr control (NR3-Scr) cells. A representative histogram is shown and average median fluorescence intensities are shown in the bar graph. Western blots were used to quantify CSL expression in the EV-Scr, EV-shCLS, NR3- Scr, and the NR3-shCSL cells. CSL expression was quantified relative to the actin protein expression (one representative WB is shown) and average CSL expression from three separate experiments is shown in the bar graph. As shown, NR3 increases FZD7 transcript and protein expression in absence of CSL expression. (n = 3, *P = £0.05, **P = £0.005, ***P = £0.0005). Color images available online at www.liebertpub.com/scd

Article Snippet: For this purpose, specific antibodies raised against FZD7 (1:100 dilution; R&D Systems clone# 151143), NOTCH1 (Cat No. A301-894A; Bethyl Laboratories), NOTCH2 (Cat No. A301-083A; Bethyl Laboratories), and NOTCH3 (Cat No. 2889; Cell Signaling) were utilized.

Techniques: Expressing, Transduction, Construct, Binding Assay, Control, Positive Control, Plasmid Preparation, Flow Cytometry, Knockdown, Western Blot

Journal: The American Journal of Pathology

Article Title: Genotypic and Phenotypic Characterization of Side Population of Gastric Cancer Cell Lines

doi: 10.1016/j.ajpath.2010.12.043

Figure Lengend Snippet: Expression of ADAM17 and FZD7 in nonneoplastic gastric tissue. A, B, C, and G: Expression of ADAM17. D, E, F, and H: Expression of FZD7. Foveolar epithelium did not express ADAM17 (B) or FZD7 (E). ADAM17 was observed in a few scattered cells of the gastric glands (C). FZD7 was expressed in inflammatory cells of the lamina propria (E), whereas the gland epithelium was immunonegative (F). ADAM17 (G) and FZD7 (H) were also expressed in bottom cells of the intestinal metaplasia. Hematoxylin counterstain. Original magnification: ×100 (A, D, G, and H); ×400 (B, C, E, and F).

Article Snippet: Immunohistochemistry Immunostaining was performed using a polyclonal antibody directed against ADAM17 (diluted 1:200; Sigma-Aldrich Corp.) and a polyclonal antibody directed against FZD7 (1:800; Aviva Systems Biology Corp., San Diego, CA).

Techniques: Expressing

Journal: The American Journal of Pathology

Article Title: Genotypic and Phenotypic Characterization of Side Population of Gastric Cancer Cell Lines

doi: 10.1016/j.ajpath.2010.12.043

Figure Lengend Snippet: Expression of ADAM17 and FZD7 in gastric cancer tissue. ADAM17 and FZD7 are expressed in intestinal-type gastric cancer cells. All images are from the same case. H&E (A), pan-cytokeratin (B), ADAM17 (C), and FZD7 (D). Hematoxylin counterstain. Original magnification: ×600 (A–D).

Article Snippet: Immunohistochemistry Immunostaining was performed using a polyclonal antibody directed against ADAM17 (diluted 1:200; Sigma-Aldrich Corp.) and a polyclonal antibody directed against FZD7 (1:800; Aviva Systems Biology Corp., San Diego, CA).

Techniques: Expressing

Journal: The American Journal of Pathology

Article Title: Genotypic and Phenotypic Characterization of Side Population of Gastric Cancer Cell Lines

doi: 10.1016/j.ajpath.2010.12.043

Figure Lengend Snippet: Correlation of Expression of ADAM17 and FZD7 With Various Clinicopathologic Patient Characteristics

Article Snippet: Immunohistochemistry Immunostaining was performed using a polyclonal antibody directed against ADAM17 (diluted 1:200; Sigma-Aldrich Corp.) and a polyclonal antibody directed against FZD7 (1:800; Aviva Systems Biology Corp., San Diego, CA).

Techniques: Expressing

Journal: The American Journal of Pathology

Article Title: Genotypic and Phenotypic Characterization of Side Population of Gastric Cancer Cell Lines

doi: 10.1016/j.ajpath.2010.12.043

Figure Lengend Snippet: Patient Survival Related to Clinicopathologic Patient Characteristics

Article Snippet: Immunohistochemistry Immunostaining was performed using a polyclonal antibody directed against ADAM17 (diluted 1:200; Sigma-Aldrich Corp.) and a polyclonal antibody directed against FZD7 (1:800; Aviva Systems Biology Corp., San Diego, CA).

Techniques:

Journal: The American Journal of Pathology

Article Title: Genotypic and Phenotypic Characterization of Side Population of Gastric Cancer Cell Lines

doi: 10.1016/j.ajpath.2010.12.043

Figure Lengend Snippet: Overall survival in dependence of FZD7-expression. Kaplan-Meier curves depict overall survival in dependence of FZD7-expression in gastric cancer. The P value was calculated using the log-rank test.

Article Snippet: Immunohistochemistry Immunostaining was performed using a polyclonal antibody directed against ADAM17 (diluted 1:200; Sigma-Aldrich Corp.) and a polyclonal antibody directed against FZD7 (1:800; Aviva Systems Biology Corp., San Diego, CA).

Techniques: Expressing

Journal: The American Journal of Pathology

Article Title: Genotypic and Phenotypic Characterization of Side Population of Gastric Cancer Cell Lines

doi: 10.1016/j.ajpath.2010.12.043

Figure Lengend Snippet: mRNA expression of ADAM17 and FZD7 in gastric tissue. A: ADAM17 mRNA is highly significantly up-regulated in gastric carcinoma compared with nonneoplastic mucosa (P < 0.001). B: FZD7 mRNA was slightly but not significantly increased in gastric cancer tissue compared with nonneoplastic mucosa (P = 0.448).

Article Snippet: Immunohistochemistry Immunostaining was performed using a polyclonal antibody directed against ADAM17 (diluted 1:200; Sigma-Aldrich Corp.) and a polyclonal antibody directed against FZD7 (1:800; Aviva Systems Biology Corp., San Diego, CA).

Techniques: Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: Schematic representation of the FZD7 protein structure and its localization within the cell membrane. Four peptides were selected for antibody generation; clones 288.1, 288.2, and 288.5 derived from peptide 4 were subsequently chosen for detailed analysis. Figure created with BioRender.com.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Membrane, Clone Assay, Derivative Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: Binding of FZD7 antibody to Wilms tumor cells, expression analysis of different WT donors and effect on cell proliferation. (A) Percentage of Frizzle 7 (FZD7)-expressing cells in culture. A representative flow cytometry analysis is shown for primary Wilms tumor (WT)-derived cells (upper) and WT-xenograft derived cells (lower). (B) A summarizing bar graph of FZD7 expression in WT cells from different donors used for in vitro experiments. (C) Effect of secreted antibodies on WT cells: Relative percentage of viable cells following 48 h of treatment with the different clones. The spent medium served as a control. The numbers indicate the hybridoma clone from which the spent medium was derived; *p < 0.05, **p < 0.01. The results are shown as the mean ± SEM of at least three different experiments for each antibody.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Binding Assay, Wilms Tumor Assay, Expressing, Flow Cytometry, Derivative Assay, In Vitro, Clone Assay, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: Binding characterization of FZD7 -288.1 in Immune Precipitation assays, Western blot analysis, FACS and Immunohistochemistry. (A) αFZD7-288.1 but not 288.5 immunoprecipitated the Frizzled 7 (FZD7) protein. A protein immunoprecipitation (IP) assay was used to validate the antibody-specific interaction with the FZD7 receptor. SK-MEL28 total cell lysates (FZD7 Input) and FZD7-bound fractions (FZD7 IP) were assayed via Western blot analysis. For detection, a commercial anti-FZD7 antibody (Millipore) was used. Nonspecific antibody and mouse IgG served as negative controls, and β-actin was used as a loading control. A representative blot of three separate experiments is shown. (B) Specific binding to the FZD7 receptor was examined in HEK293T cells overexpressing full-length FZD7. The cell extracts were probed with different concentrations of αFZD7-288.1. Specific binding of the Ab to FZD7-expressing cells was observed. (C) Probing of cell lysates from sh-FZD7-Wilms tumor (WT) cells with αFZD7-288.1 revealed the downregulation of FZD7, similar to the effect of a commercial αFZD7 antibody, demonstrating its specificity. β-Actin was used as a loading control. (D) Representative flow cytometry analysis of FZD7 expression in cultured WT cells stained with both αFZD7-288.1 and a rat anti-FZD7 APC-conjugated antibody. Cells stained positive for αFZD7-288.1 were mostly observed as a subpopulation within the cells detected by RαFZD7-APC. The data are presented in a dot plot graph showing FZD7 staining in red and isotype control staining (negative control) in gray.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Binding Assay, Western Blot, Immunohistochemistry, Immunoprecipitation, Control, Expressing, Wilms Tumor Assay, Flow Cytometry, Cell Culture, Staining, Negative Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: The αFZD7-288.1 antibody inhibits β-catenin translocating Wnt signaling in Wilms tumor cells. (A) Western blot (WB) analysis demonstrating the inhibition of active β-catenin and decreased levels of Fizzled 7 (FZD7). (B) following treatment with 5 μg/mL αFZD7-288.1 for 48 h in Wilms tumor (WT) cells. A nonspecific Ab was used as a negative control; β-actin was used as a loading control. (C) β-catenin immunofluorescence staining of untreated WT and αFZD7-288.1-treated cells: Double-staining with DAPI and cytokeratin (first panel) and with cytokeratin and β-catenin (second panel). β-catenin staining revealed strong expression and a greater percentage of cells with nuclear localization of β-catenin in the control untreated cells (scale bar represents 50 µM) X70 magnification of representative cells (indicated by arrows) from each group (third panel) and double-staining with DAPI and β-catenin (forth panel). (D–F) Real-time PCR gene expression analysis of WT cells: The expression of Wnt transducers [ (D) ; FZD7, β-catenin], Wnt pathway target genes [ (E) ; AXIN2, CCND1, MYC-C], and Wnt pathway inhibitors [ (F) ; DKK1, sFRP1] was also measured. The results showed reduced mRNA expression of Wnt pathway-related genes. The data are shown as the mean ± S.E.M. of 5 separate experiments; *p < 0.05; **p < 0.01.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Wilms Tumor Assay, Western Blot, Inhibition, Negative Control, Control, Immunofluorescence, Staining, Double Staining, Expressing, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: The αFZD7-288.1 antibody reduces proliferation in Wilms tumors cells and induces cell death. (A) Inhibition of the Wnt pathway by αFZD7-288.1 treatment induced changes in cell kinetics, as demonstrated by the longer doubling time of antibody treated Wilms tumor (WT) cells compared to control treated WT cells and (B) the significantly lower growth rate (defined as the number of cell divisions per day), *p < 0.05; n = 4. Reduced proliferation in cells treated with αFZD7-288.1 was also observed by MTS assay in (C) and (D) Quantification of Ki67 staining was performed by counting the number of Ki67-positive nuclei in five high-power fields (HPF) per condition, using representative regions from each sample. Reduced nuclear Ki67 expression (red, lower panels) compared to that in control untreated cells (magnification ×40 scale bars = 100 μm; magnification ×20 scale bars = 50 µm). (E) Induction of cell death in WT cells is shown in a representative analysis for annexin V staining, which demonstrated a marked increase in the percentage of apoptotic and preapoptotic cells. (F) Percentage of dead cells calculated using trypan blue staining of WT cells indicating significantly greater cell death in treated WT cells than in control cells; *p < 0.05; n = 4.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Inhibition, Wilms Tumor Assay, Control, MTS Assay, Staining, Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: The migration and sphere formation of Wilms tumor cells is reduced after treatment with αFZD7-288.1 antibody. (A) Scratch assay demonstrating the effects of αFZD7-288.1 on the ability of cells to migrate through a scrape in the cell monolayer. Wilms tumor (WT)-treated cells demonstrated a lower migration capacity than control cells, as shown in the representative images. Calculation of the area of migrating cells (on the right) showed that the cells had a significantly reduced migration capacity following treatment; *p < 0.05; n = 3. (B) A sphere formation assay was performed. The number and size of the spheres formed by the treated cells were significantly lower than those formed by the untreated cells. The data are presented as the means ± SEMs of at least three experiments performed in triplicate (*p < 0.05) On the left, representative phase-contrast images of spheres formed by treated and untreated WT cells showing markedly fewer condensed spheres in the treated cells (scale bars = 100 μm, magnification ×10).

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Migration, Wilms Tumor Assay, Wound Healing Assay, Control, Tube Formation Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: αFZD7-288.1 strongly co-localizes with NCAM1 in WT, and treatment with αFZD7-288.1 reduces WT CSC markers. (A) Immunofluorescence of staining of cultured WT cells using αFZD7-288.1 and NCAM1 and co-staining (αFZD7-red, αNCAM1 green and DAPI blue) microscopy revealed overlapping signals as a merged color (yellow), indicating membrane spatial proximity. Slides were imaged using Olympus IX83 fluorescent microscope and Olympus DP80 camera. Image processing was done using Olympus OlyVia software. (B) The expression of the Cancer stem cell marker genes NCAM1, CITED, SIX2 and ALDH1A1 decreased in the antibody αFZD7-288.1 treated WT derived cells (n = 3); *p < 0.05; **p < 0.01.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Immunofluorescence, Staining, Cell Culture, Microscopy, Membrane, Software, Expressing, Marker, Derivative Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: The antitumor activity of αFZD7-288.1 as demonstrated in changes in tumor volumes. (A) Flow cytometry histogram evaluating Frizzled 7 (FZD7) expression in Wilms tumor (WT) tissue for use in vivo . (B) Workflow diagram illustrating the in vivo experimental design. (C) Mouse weights during treatment. Paclitaxel (PTX)-treated mice suffered from toxic side effects and significant weight loss following treatment; **p < 0.01; ***p < 0.005. (D) Antitumor activity: Mice were treated intravenously with normal saline, αFZD7-288.1 (10 mg/kg) or paclitaxel (15 mg/kg). Tumor volume, presented in mm 3 , was assessed every 2–3 days using an external electronic caliper and calculated by the modified ellipsoid formula V = 0.52 × (length × width 2 ). Like PTX-treated tumors, tumors treated with αFZD7-288.1 exhibited growth inhibition and even tumor shrinkage, while saline-treated tumors showed rapid tumor growth; *p < 0.05; **p < 0.01. (E) Representative images on day 10 of the experiment showing mice with large saline-treated tumors (right) and unnoticeable αFZD7-288.1-treated tumors (left). (F) Waterfall graph showing the changes in the individual tumor volumes with respect to the initial tumor size. (G) FZD7 expression in tumors treated with αFZD7-288.1 was significantly altered by 0.445-fold; *p < 0.05; n = 3 per group. (H) The expression of the canonical Wnt pathway genes FZD7, β-CATENIN, AXIN2, CCND1, C-MYC, DKK1, and sFRP1 decreased in the treated tumors (n = 4); *p < 0.05; **p < 0.01; ***p < 0.005.

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Activity Assay, Flow Cytometry, Expressing, Wilms Tumor Assay, In Vivo, Saline, Modification, Inhibition

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel Frizzled 7 antibody disrupts the Wnt pathway and inhibits Wilms tumor growth

doi: 10.3389/fbioe.2025.1641137

Figure Lengend Snippet: The antitumor activity of αFZD7-288.1 as demonstrated by proliferative markers in Immunohistochemical staining. (A) Immunohistochemical staining for Ki67 and Caspase3 in representative tumors from the different experimental groups; saline, paclitaxel (PTX) and αFZD7-288.1 (type A, representing reduced-volume tumors; and type B, representing tumors with growth inhibition compared to saline-treated tumors). The scale bars represent 50 μm at a magnification of ×20. Compared with those of saline- or PTX-treated tumors, αFZD7-288.1-treated tumors exhibited reduced proliferation. An increase in apoptosis was observed in αFZD7-288.1- and PTX-treated tumors (B) .

Article Snippet: The cells were stained with an isotype control (R and D Systems Cat# IC006A, RRID:AB_357254) or a primary antibody against FZD7 (R and D Systems Cat# FAB1981A, RRID:AB_2232278).

Techniques: Activity Assay, Immunohistochemical staining, Staining, Saline, Inhibition

Journal: Scientific reports

Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis.

doi: 10.1038/s41598-025-90803-9

Figure Lengend Snippet: Fig. 2. (A, C, E, G, I) FZD7 is significantly up-regulated in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets. (B, D, F, H, J) The receiver-operating characteristic (ROC) curve and the area under the curve (AUC) curve of FZD7 in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets.

Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

Techniques:

Journal: Scientific reports

Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis.

doi: 10.1038/s41598-025-90803-9

Figure Lengend Snippet: Fig. 3. GO and KEGG pathways for the overlapping FZD7 positively associated genes in GSE11691, GSE7305, GSE5108, GSE23339 and GSE25628 datasets.

Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

Techniques:

Journal: Scientific reports

Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis.

doi: 10.1038/s41598-025-90803-9

Figure Lengend Snippet: Fig. 4. (A, C) The infiltration of 22 immune cells in EMS of GSE11691dataset. (B) The relationship between FZD7 and 22 immune cells in GSE11691 dataset. (D) The expression of FZD7 in GSE19834. (E) Immunofluorescence images staining with CD206 in Vehicle group and F7H group.

Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Scientific reports

Article Title: Identification of FZD7 as a potential ferroptosis-related diagnostic gene in endometriosis by bioinformatics analysis.

doi: 10.1038/s41598-025-90803-9

Figure Lengend Snippet: Fig. 5. (A-C) The expression of FZD7 in ectopic (Ec) endometrial tissues, eutopic (Eu) endometrial tissues and normal endometrial (NE) tissues. **p < 0.01, ***p < 0.001, ns, no significant difference by one-way analysis of variance (ANOVA) test.

Article Snippet: The membrane was blocked with 5% skimmed milk powder solution for 2 h, then incubated with primary antibody against FZD7 (16974-1-AP, Proteintech, China) overnight at 4 °C, followed by incubation with secondary antibody for 2 h at room temperature.

Techniques: Expressing