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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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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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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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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 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