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wnt 3a  (R&D Systems)


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    Structured Review

    R&D Systems wnt 3a
    Wnt 3a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+wnt+3a/Recombinant+Human+Wnt-3a+Protein/pm41933211-578-43-45
    Average 96 stars, based on 524 article reviews
    wnt 3a - by Bioz Stars, 2026-10
    96/100 stars

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    Recombinant:

    Article Title: Inhibition of PORCN Blocks Wnt Signaling to Attenuate Progression of Oral Carcinogenesis
    Article Snippet: Purpose: Oral squamous cell carcinoma (OSCC) is commonly preceded by potentially malignant lesions, referred to as oral dysplasia.. We recently reported that oral dysplasia is associated with aberrant activation of the Wnt/b-catenin pathway, due to overexpression of Wnt ligands in a Porcupine (PORCN)-dependent manner.. Pharmacologic inhibition of PORCN precludesWnt secretion and has been proposed as a potential therapeutic approach to treat established cancers.

    Article Title: Integrin-linked kinase-frizzled 7 interaction maintains cancer stem cells to drive platinum resistance in ovarian cancer
    Article Snippet: .. Recombinant human Wnt-3A was purchased from R&D Systems (Minneapolis, MN). .. ILK inhibitor, cpd-22 was purchased from Millipore-Sigma (Burlington, MA, USA).

    Article Title: Machine learning on large scale perturbation screens for SARS-CoV-2 host factors identifies β-catenin/CBP inhibitor PRI-724 as a potent antiviral.
    Article Snippet: Compounds were resuspended in DMSO (Carl Roth; Karlsruhe, Germany) and aliquots were stored at −80°C. .. Recombinant human Wnt-3a (rhWnt-3a; #5036-WN, R&D Systems; Minneapolis, Minnesota, United States) was resuspended according to manufacturer’s instructions and stored at −80°C. ..

    Article Title: Machine learning on large scale perturbation screens for SARS-CoV-2 host factors identifies β-catenin/CBP inhibitor PRI-724 as a potent antiviral
    Article Snippet: Compounds were resuspended in DMSO (Carl Roth; Karlsruhe, Germany) and aliquots were stored at −80°C. .. Recombinant human Wnt-3a (rhWnt-3a; #5036-WN, R&D Systems; Minneapolis, Minnesota, United States) was resuspended according to manufacturer’s instructions and stored at −80°C. ..

    Article Title: Integrin-linked kinase-frizzled 7 interaction maintains cancer stem cells to drive platinum resistance in ovarian cancer
    Article Snippet: .. Recombinant human Wnt-3A was purchased from R&D Systems (Minneapolis, MN). .. ILK inhibitor, cpd-22 was purchased from Millipore-Sigma (Burlington, MA, USA).

    Article Title: CRISPR/Cas9-mediated gene correction of Wilson disease H1069Q point mutation in patient-specific induced pluripotent stem cells.
    Article Snippet: .. Briefly, cells were incubated in DMEM/F12 (Gibco, Thermo Fisher Scientific) with increasing KnockOut SR Xenofree medium (KSR; Gibco, Thermo Fisher Scientific) concentrations, enriched with 100 ng/μl recombinant Activin-A (R&D Systems, Minneapolis, MN, USA), 100 ng/μl FGF2 (PeproTech, Thermo Fisher Scientific), and 10 μg/ml recombinant human Wnt-3a (R&D Systems) for 3 days (day 1 to 3). .. For the next 8 days (days 4 to 11), cells were cultivated in DMEM/ F12 supplemented with 10% KSR, 1 mM NEAA, 1 mM L-glutamine (Life Technologies), 1% dimethyl sulfoxide (PanReacAppliChem, Darmstadt, Germany), and 100 ng/μl hepatocyte growth factor (PeproTech, Thermo Fisher Scientific).

    other:

    Article Title: Insights into the Structure and Dynamics of Human Wnt Signal Transduction
    Article Snippet: For iPSCs, Both Piggyback and Donor plasmids were chemically transfected when cells reached 30% confluency using Lipofectamine Stem Transfection Reagent (manufactures protocol).

    Incubation:

    Article Title: CRISPR/Cas9-mediated gene correction of Wilson disease H1069Q point mutation in patient-specific induced pluripotent stem cells.
    Article Snippet: .. Briefly, cells were incubated in DMEM/F12 (Gibco, Thermo Fisher Scientific) with increasing KnockOut SR Xenofree medium (KSR; Gibco, Thermo Fisher Scientific) concentrations, enriched with 100 ng/μl recombinant Activin-A (R&D Systems, Minneapolis, MN, USA), 100 ng/μl FGF2 (PeproTech, Thermo Fisher Scientific), and 10 μg/ml recombinant human Wnt-3a (R&D Systems) for 3 days (day 1 to 3). .. For the next 8 days (days 4 to 11), cells were cultivated in DMEM/ F12 supplemented with 10% KSR, 1 mM NEAA, 1 mM L-glutamine (Life Technologies), 1% dimethyl sulfoxide (PanReacAppliChem, Darmstadt, Germany), and 100 ng/μl hepatocyte growth factor (PeproTech, Thermo Fisher Scientific).

    Knock-Out:

    Article Title: CRISPR/Cas9-mediated gene correction of Wilson disease H1069Q point mutation in patient-specific induced pluripotent stem cells.
    Article Snippet: .. Briefly, cells were incubated in DMEM/F12 (Gibco, Thermo Fisher Scientific) with increasing KnockOut SR Xenofree medium (KSR; Gibco, Thermo Fisher Scientific) concentrations, enriched with 100 ng/μl recombinant Activin-A (R&D Systems, Minneapolis, MN, USA), 100 ng/μl FGF2 (PeproTech, Thermo Fisher Scientific), and 10 μg/ml recombinant human Wnt-3a (R&D Systems) for 3 days (day 1 to 3). .. For the next 8 days (days 4 to 11), cells were cultivated in DMEM/ F12 supplemented with 10% KSR, 1 mM NEAA, 1 mM L-glutamine (Life Technologies), 1% dimethyl sulfoxide (PanReacAppliChem, Darmstadt, Germany), and 100 ng/μl hepatocyte growth factor (PeproTech, Thermo Fisher Scientific).



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    R&D Systems wnt 3a
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    Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K <t>,</t> <t>dual-luciferase</t> assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant WNT3A protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.
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    R&D Systems human wnt3a protein
    Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K , dual-luciferase assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant <t>WNT3A</t> protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.
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    Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K , dual-luciferase assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant <t>WNT3A</t> protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.
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    β-lactamase activity concentration-response to compounds in LEF/TCF-bla FreeStyle™ 293F cell in 1536-well format. (A) Stimulation concentration response curve of <t>human</t> <t>WNT-3A</t> <t>(hWNT-3A).</t> (B) Stimulation concentration response curves of the Wnt potentiator R-Spondin2 (hRS) in combination with different concentrations of hWNT-3A (hRS alone, 75, 150, and 300 ng/mL). (C) Curves showing the mild effect of imatinib mesylate on cell viability and its inhibitory effect on stimulator-induced β-lactamase activity, confirming assay responsiveness to known Wnt pathway antagonists.
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    β-lactamase activity concentration-response to compounds in LEF/TCF-bla FreeStyle™ 293F cell in 1536-well format. (A) Stimulation concentration response curve of <t>human</t> <t>WNT-3A</t> <t>(hWNT-3A).</t> (B) Stimulation concentration response curves of the Wnt potentiator R-Spondin2 (hRS) in combination with different concentrations of hWNT-3A (hRS alone, 75, 150, and 300 ng/mL). (C) Curves showing the mild effect of imatinib mesylate on cell viability and its inhibitory effect on stimulator-induced β-lactamase activity, confirming assay responsiveness to known Wnt pathway antagonists.
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    Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K , dual-luciferase assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant WNT3A protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.

    Journal: The Journal of Biological Chemistry

    Article Title: IFIT3-DVL interaction promotes malignant progression of lung squamous cell carcinoma and large-cell lung carcinoma via canonical WNT signaling

    doi: 10.1016/j.jbc.2026.111368

    Figure Lengend Snippet: Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K , dual-luciferase assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant WNT3A protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.

    Article Snippet: For analysis of WNT3A response, cells were treated with recombinant human WNT3A protein (100 ng/ml, R&D systems, Cat# 5036-WN-500/CF) for 12 h prior to dual-luciferase assay or Western blot analysis.

    Techniques: Western Blot, Control, Phospho-proteomics, Real-time Polymerase Chain Reaction, Plasmid Preparation, Expressing, Transfection, Negative Control, Knockdown, Luciferase, Recombinant, Positive Control, Two Tailed Test

    Effect of IFIT3 on canonical WNT signaling. A , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in A . Vec, vector, is the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. After transfection with IFIT3 expression plasmid, cells were treated for 24 h with dimethyl sulfoxide (10 μM; IFIT3+ dimethyl sulfoxide group) or XAV-939 (10 μM; IFIT3+XAV-939 group). B , dual-luciferase assay. TOPFLASH/FOPFLASH was used to measure for β-catenin/TCF mediated transcriptional activity. C , nuclear and Cytoplasmic Protein Extraction Assay analysis. Cyto, cytoplasm; Nuc, nucleus; Lamin B1 was a reference nuclear protein, and α-tubulin was a positive control for the cytoplasm. Quantitative analysis of band intensities is provided in B . D , immunofluorescence analysis (the scale bar represents 10 μm). Quantification of average fluorescence intensity was provided. E , representative immunohistochemistry images for IFIT3 and β-catenin in LUSC and LCLC (magnification: × 400; the scale bar represents 50 μm). F , Kaplan-Meier survival curve analyzing the different group in cancerous samples from LUSC/LCLC patients. a, the group with cytoplasmic IFIT3 and nuclear β-catenin coexpression (16 cases); b, the group with IFIT3 and β-catenin coexpression in the cytoplasm (56 cases); c. others (110 cases). Log-rank test was used for survival comparison. Error bars indicated mean ± SEM (n = 3). Data were analyzed by one-way ANOVA with Tukey’s post hoc test for ( B ), and two-tailed unpaired Student’s t test for ( D ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.

    Journal: The Journal of Biological Chemistry

    Article Title: IFIT3-DVL interaction promotes malignant progression of lung squamous cell carcinoma and large-cell lung carcinoma via canonical WNT signaling

    doi: 10.1016/j.jbc.2026.111368

    Figure Lengend Snippet: Effect of IFIT3 on canonical WNT signaling. A , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in A . Vec, vector, is the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. After transfection with IFIT3 expression plasmid, cells were treated for 24 h with dimethyl sulfoxide (10 μM; IFIT3+ dimethyl sulfoxide group) or XAV-939 (10 μM; IFIT3+XAV-939 group). B , dual-luciferase assay. TOPFLASH/FOPFLASH was used to measure for β-catenin/TCF mediated transcriptional activity. C , nuclear and Cytoplasmic Protein Extraction Assay analysis. Cyto, cytoplasm; Nuc, nucleus; Lamin B1 was a reference nuclear protein, and α-tubulin was a positive control for the cytoplasm. Quantitative analysis of band intensities is provided in B . D , immunofluorescence analysis (the scale bar represents 10 μm). Quantification of average fluorescence intensity was provided. E , representative immunohistochemistry images for IFIT3 and β-catenin in LUSC and LCLC (magnification: × 400; the scale bar represents 50 μm). F , Kaplan-Meier survival curve analyzing the different group in cancerous samples from LUSC/LCLC patients. a, the group with cytoplasmic IFIT3 and nuclear β-catenin coexpression (16 cases); b, the group with IFIT3 and β-catenin coexpression in the cytoplasm (56 cases); c. others (110 cases). Log-rank test was used for survival comparison. Error bars indicated mean ± SEM (n = 3). Data were analyzed by one-way ANOVA with Tukey’s post hoc test for ( B ), and two-tailed unpaired Student’s t test for ( D ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.

    Article Snippet: For analysis of WNT3A response, cells were treated with recombinant human WNT3A protein (100 ng/ml, R&D systems, Cat# 5036-WN-500/CF) for 12 h prior to dual-luciferase assay or Western blot analysis.

    Techniques: Western Blot, Control, Plasmid Preparation, Expressing, Transfection, Luciferase, Activity Assay, Protein Extraction, Positive Control, Immunofluorescence, Fluorescence, Immunohistochemistry, Comparison, Two Tailed Test

    IFIT3 interacts with DVL2 to promote DVL2 phosphorylation and upregulate β-catenin expression. A , co-immunoprecipitation analysis. IP, immunoprecipitation. IB, immunoblot. B , immunofluorescence assay (the scale bar represents 50 μm). Quantitative colocalization analysis of these images is presented in . C, E and F , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. p-β-catenin (Ser675), the serine 675 phosphorylation of β-catenin. ABC, active non-phospho (Ser33/37/Thr41) β-catenin. p-DVL2 (Ser143), the serine 143 phosphorylation of DVL2. p-DVL2 (Thr224), the threonine 224 phosphorylation of DVL2. D , dual-luciferase assay. TOPFLASH/FOPFLASH was measured for β-catenin/TCF mediated transcriptional activity. Cells were co-transfected with siRNA and plasmid. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-D1, siRNA-DVL1. si-D2, siRNA-DVL2. si-D3, siRNA-DVL3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. Data were analyzed by one-way ANOVA with Tukey’s post hoc test for ( D ). Error bars indicated mean ± SEM (n = 3). ∗ p < 0.05.

    Journal: The Journal of Biological Chemistry

    Article Title: IFIT3-DVL interaction promotes malignant progression of lung squamous cell carcinoma and large-cell lung carcinoma via canonical WNT signaling

    doi: 10.1016/j.jbc.2026.111368

    Figure Lengend Snippet: IFIT3 interacts with DVL2 to promote DVL2 phosphorylation and upregulate β-catenin expression. A , co-immunoprecipitation analysis. IP, immunoprecipitation. IB, immunoblot. B , immunofluorescence assay (the scale bar represents 50 μm). Quantitative colocalization analysis of these images is presented in . C, E and F , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. p-β-catenin (Ser675), the serine 675 phosphorylation of β-catenin. ABC, active non-phospho (Ser33/37/Thr41) β-catenin. p-DVL2 (Ser143), the serine 143 phosphorylation of DVL2. p-DVL2 (Thr224), the threonine 224 phosphorylation of DVL2. D , dual-luciferase assay. TOPFLASH/FOPFLASH was measured for β-catenin/TCF mediated transcriptional activity. Cells were co-transfected with siRNA and plasmid. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-D1, siRNA-DVL1. si-D2, siRNA-DVL2. si-D3, siRNA-DVL3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. Data were analyzed by one-way ANOVA with Tukey’s post hoc test for ( D ). Error bars indicated mean ± SEM (n = 3). ∗ p < 0.05.

    Article Snippet: For analysis of WNT3A response, cells were treated with recombinant human WNT3A protein (100 ng/ml, R&D systems, Cat# 5036-WN-500/CF) for 12 h prior to dual-luciferase assay or Western blot analysis.

    Techniques: Phospho-proteomics, Expressing, Immunoprecipitation, Western Blot, Immunofluorescence, Control, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Knockdown

    Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K , dual-luciferase assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant WNT3A protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.

    Journal: The Journal of Biological Chemistry

    Article Title: IFIT3-DVL interaction promotes malignant progression of lung squamous cell carcinoma and large-cell lung carcinoma via canonical WNT signaling

    doi: 10.1016/j.jbc.2026.111368

    Figure Lengend Snippet: Effect of IFIT3 on cell malignancy and canonical WNT signaling. A and B , Western blot analysis. GAPDH was an internal control. Quantitative analysis of band intensities is provided in . p-GSK-3β (Ser9), the serine nine phosphorylation of GSK-3β. C–H , quantitative real-time PCR analysis. β-actin was an internal control. Vec, vector, the control group of the IFIT3 expression plasmid. IF3, transfection of IFIT3 expression plasmid. si-IF3, siRNA-IFIT3. si-NC, siRNA-negative control, a scrambled siRNA selected as a negative control. si-IF3 (IF3), following IFIT3 knockdown with siRNA, its expression was restored by transfection with IFIT3 expression plasmid. I–K , dual-luciferase assay. IFIT3 expression plasmid or IFIT3 siRNA was transfected to LUSC/LCLC cells, with or without the treatment of recombinant WNT3A protein. (i) Negative Control: transfection of empty plasmid vector or siRNA-negative control. (ii) Positive Control: treatment of recombinant WNT3A protein. (iii) Test Group 1: transfection of IFIT3 expression plasmid or IFIT3 siRNA. (iv) Test Group 2: transfection of IFIT3 expression plasmid or IFIT3 siRNA, with the treatment of recombinant WNT3A protein. rhW3A, the cells were treated with recombinant human WNT3A protein (100 ng/ml) for 12 h. Error bars indicated mean ± SEM (n = 3). Data were analyzed by two-tailed unpaired Student’s t test for ( C , D , F and G ), and one-way ANOVA with Tukey’s post hoc test for ( E , H–K ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. LCLC, large-cell lung carcinoma; LUSC, lung squamous cell carcinoma.

    Article Snippet: For analysis of WNT3A response, cells were treated with recombinant human WNT3A protein (100 ng/ml, R&D systems, Cat# 5036-WN-500/CF) for 12 h prior to dual-luciferase assay or Western blot analysis.

    Techniques: Western Blot, Control, Phospho-proteomics, Real-time Polymerase Chain Reaction, Plasmid Preparation, Expressing, Transfection, Negative Control, Knockdown, Luciferase, Recombinant, Positive Control, Two Tailed Test

    β-lactamase activity concentration-response to compounds in LEF/TCF-bla FreeStyle™ 293F cell in 1536-well format. (A) Stimulation concentration response curve of human WNT-3A (hWNT-3A). (B) Stimulation concentration response curves of the Wnt potentiator R-Spondin2 (hRS) in combination with different concentrations of hWNT-3A (hRS alone, 75, 150, and 300 ng/mL). (C) Curves showing the mild effect of imatinib mesylate on cell viability and its inhibitory effect on stimulator-induced β-lactamase activity, confirming assay responsiveness to known Wnt pathway antagonists.

    Journal: Frontiers in Pharmacology

    Article Title: Quantitative high-throughput screening identifies small-molecule modulators of Wnt/β-catenin signaling

    doi: 10.3389/fphar.2026.1788041

    Figure Lengend Snippet: β-lactamase activity concentration-response to compounds in LEF/TCF-bla FreeStyle™ 293F cell in 1536-well format. (A) Stimulation concentration response curve of human WNT-3A (hWNT-3A). (B) Stimulation concentration response curves of the Wnt potentiator R-Spondin2 (hRS) in combination with different concentrations of hWNT-3A (hRS alone, 75, 150, and 300 ng/mL). (C) Curves showing the mild effect of imatinib mesylate on cell viability and its inhibitory effect on stimulator-induced β-lactamase activity, confirming assay responsiveness to known Wnt pathway antagonists.

    Article Snippet: Recombinant human WNT-3A (hWNT-3A) and R-spondin 2 (hRS) were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Activity Assay, Concentration Assay

    Western blot confirmation of identified inhibitors. (A) Representative Western blot analysis showing the effects of each compound on β-catenin protein levels. Treatment with the stimulator (DMSO +) markedly increased both non-phosphorylated (active) and total β-catenin compared with the DMSO control (DMSO -). All known inhibitors attenuated the stimulator-induced increases. (B) Quantitative densitometry of non-phosphorylated and total β-catenin from three independent experiments, normalized to the housekeeping protein GAPDH. Data are presented as mean ± SD. Statistical significances relative to the DMSO + control is indicated above bars (*p < 0.05; **p < 0.01; ***p < 0.001). “+” denotes the addition of stimulator; “-” denotes its absence. The stimulator consisted of 75 ng/mL hWNT-3A and 95 ng/mL hRS.

    Journal: Frontiers in Pharmacology

    Article Title: Quantitative high-throughput screening identifies small-molecule modulators of Wnt/β-catenin signaling

    doi: 10.3389/fphar.2026.1788041

    Figure Lengend Snippet: Western blot confirmation of identified inhibitors. (A) Representative Western blot analysis showing the effects of each compound on β-catenin protein levels. Treatment with the stimulator (DMSO +) markedly increased both non-phosphorylated (active) and total β-catenin compared with the DMSO control (DMSO -). All known inhibitors attenuated the stimulator-induced increases. (B) Quantitative densitometry of non-phosphorylated and total β-catenin from three independent experiments, normalized to the housekeeping protein GAPDH. Data are presented as mean ± SD. Statistical significances relative to the DMSO + control is indicated above bars (*p < 0.05; **p < 0.01; ***p < 0.001). “+” denotes the addition of stimulator; “-” denotes its absence. The stimulator consisted of 75 ng/mL hWNT-3A and 95 ng/mL hRS.

    Article Snippet: Recombinant human WNT-3A (hWNT-3A) and R-spondin 2 (hRS) were obtained from R&D Systems (Minneapolis, MN, USA).

    Techniques: Western Blot, Control