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Cusabio immunoassay elisa kit
Immunoassay Elisa Kit, supplied by Cusabio, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wnt4+elisa+kits/Human+Protein+Wnt-4(WNT4)+ELISA+kit/pm34898058-59-13-22
Average 92 stars, based on 4 article reviews

immunoassay elisa kit - by Bioz Stars, 2026-09

92/100 stars

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Enzyme-linked Immunosorbent Assay:

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction
Article Snippet: .. Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions. .. C57BL/6 male mice (8–10 weeks, Charles River Laboratories) were used to build MI model.

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Effects of LPS on inflammation and apoptosis in HDPCs. The HDPCs were treated with LPS for 24 h. (A) The morphology of HDPCs was observed under an optical microscope. Scale bar, 100 µm. (B) Expressions of vimentin and keratin in HDPCs were measured by immunofluorescence. Scale bar, 100 µm/30 µm. (C) The expressions of inflammatory factors and (D) apoptosis-related proteins were detected by western blotting. (E) Flow cytometry was applied to measure cell apoptosis. (F) Reverse transcription-quantitative PCR and (G) western blotting was used to detect the expression of Wnt4. Data represent the mean values ± standard deviation of three experiments. ** P<0.01. LPS, lipopolysaccharide; HDPCs, human dental pulp cells.

Journal: Experimental and Therapeutic Medicine

Article Title: Wnt4 prevents apoptosis and inflammation of dental pulp cells induced by LPS by inhibiting the IKK/NF‑κB pathway

doi: 10.3892/etm.2022.11774

Figure Lengend Snippet: Effects of LPS on inflammation and apoptosis in HDPCs. The HDPCs were treated with LPS for 24 h. (A) The morphology of HDPCs was observed under an optical microscope. Scale bar, 100 µm. (B) Expressions of vimentin and keratin in HDPCs were measured by immunofluorescence. Scale bar, 100 µm/30 µm. (C) The expressions of inflammatory factors and (D) apoptosis-related proteins were detected by western blotting. (E) Flow cytometry was applied to measure cell apoptosis. (F) Reverse transcription-quantitative PCR and (G) western blotting was used to detect the expression of Wnt4. Data represent the mean values ± standard deviation of three experiments. ** P<0.01. LPS, lipopolysaccharide; HDPCs, human dental pulp cells.

Article Snippet: The membrane was blocked with 5% skimmed milk at room temperature for 60 min and incubated with following primary antibodies: IL-8 (cat. no. ab289967; 1:1,200; Abcam), IL-6 (cat. no. ab233706; 1:1,000; Abcam), TNF-α (cat. no. ab183218; 1:1,000; Abcam), IL-1β (cat. no. ab254360; 1:1,000; Abcam), cleaved-caspase-3 (cat. no. ab32042; 1:800; Abcam), caspase-3 (cat. no. ab32351; 1:3,000; Abcam), Bax (cat. no. 2774S; 1:1,000; CST), Bcl-2 (cat. no. 15071; 1:1,000; CST), Wnt4 (cat. no. ab277798; 1:800, Abcam), p-IKK2 (cat. no. 2694; 1:1,000; CST), IKK (cat. no. ab124957; 1:1,000; Abcam), p-p65 (cat. no. 3033; 1:1,000; CST), p65 (cat. no. 8242; 1:1,000; CST), p-IκBα (cat. no. 2859; 1:1,000; CST), IκBα (cat. no. 9242; 1:1,000; CST), β-actin (cat. no. 4970; 1:5,000; CST), GAPDH (cat. no. 5174; 1:5,000; CST) at 4 ̊C overnight.

Techniques: Microscopy, Immunofluorescence, Western Blot, Flow Cytometry, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

Overexpression of Wnt4 inhibits inflammation and apoptosis in LPS-induced HDPCs. pcDNA-Wnt4 or pcDNA-NC plasmid was transfected into LPS-HDPCs. (A) Reverse transcription-quantitative PCR and (B) western blotting were applied to detect the expression of Wnt4 after transfection with pcDNA-Wnt4 or pcDNA-NC in LPS-HDPCs. The expressions of (C) inflammatory factors and (D) apoptosis-related proteins were detected by western blotting. (E) Flow cytometry was applied to measure cell apoptosis. Data represent the mean values ± standard deviation of three experiments. ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; OE-Wnt4, overexpression of Wnt4; OE-NC, negative control; p-, phosphorylated.

Journal: Experimental and Therapeutic Medicine

Article Title: Wnt4 prevents apoptosis and inflammation of dental pulp cells induced by LPS by inhibiting the IKK/NF‑κB pathway

doi: 10.3892/etm.2022.11774

Figure Lengend Snippet: Overexpression of Wnt4 inhibits inflammation and apoptosis in LPS-induced HDPCs. pcDNA-Wnt4 or pcDNA-NC plasmid was transfected into LPS-HDPCs. (A) Reverse transcription-quantitative PCR and (B) western blotting were applied to detect the expression of Wnt4 after transfection with pcDNA-Wnt4 or pcDNA-NC in LPS-HDPCs. The expressions of (C) inflammatory factors and (D) apoptosis-related proteins were detected by western blotting. (E) Flow cytometry was applied to measure cell apoptosis. Data represent the mean values ± standard deviation of three experiments. ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; OE-Wnt4, overexpression of Wnt4; OE-NC, negative control; p-, phosphorylated.

Techniques: Over Expression, Plasmid Preparation, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Flow Cytometry, Standard Deviation, Negative Control

Wnt4 suppresses the activation of IKK/NF-κB pathway in LPS-induced HDPCs. (A) Western blotting and (B) immunofluorescence staining were used to analyze the activation of IKK/NF-κB pathway. Data represent the mean values ± standard deviation of three experiments. ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; p-, phosphorylated; OE-Wnt4, overexpression of Wnt4; OE-NC, negative control.

Journal: Experimental and Therapeutic Medicine

Article Title: Wnt4 prevents apoptosis and inflammation of dental pulp cells induced by LPS by inhibiting the IKK/NF‑κB pathway

doi: 10.3892/etm.2022.11774

Figure Lengend Snippet: Wnt4 suppresses the activation of IKK/NF-κB pathway in LPS-induced HDPCs. (A) Western blotting and (B) immunofluorescence staining were used to analyze the activation of IKK/NF-κB pathway. Data represent the mean values ± standard deviation of three experiments. ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; p-, phosphorylated; OE-Wnt4, overexpression of Wnt4; OE-NC, negative control.

Techniques: Activation Assay, Western Blot, Immunofluorescence, Staining, Standard Deviation, Over Expression, Negative Control

Wnt4 inhibits inflammation and apoptosis through the IKK/NF-κB pathway. pcDNA-Wnt4 or negative control and pCMV-IKK2EE or negative control plasmids were transfected in LPS-HDPCs. (A) Western blotting and (B) immunofluorescence staining were used to measure protein levels of the IKK/NF-κB pathway. The expressions of (C) inflammatory factors and (D) apoptosis-related proteins were detected by western blot. (E) Flow cytometry was applied to measure cell apoptosis. Data represent the mean values ± standard deviation of three experiments. ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; OE-Wnt4, overexpression of Wnt4, OE-NC, negative control.

Journal: Experimental and Therapeutic Medicine

Article Title: Wnt4 prevents apoptosis and inflammation of dental pulp cells induced by LPS by inhibiting the IKK/NF‑κB pathway

doi: 10.3892/etm.2022.11774

Figure Lengend Snippet: Wnt4 inhibits inflammation and apoptosis through the IKK/NF-κB pathway. pcDNA-Wnt4 or negative control and pCMV-IKK2EE or negative control plasmids were transfected in LPS-HDPCs. (A) Western blotting and (B) immunofluorescence staining were used to measure protein levels of the IKK/NF-κB pathway. The expressions of (C) inflammatory factors and (D) apoptosis-related proteins were detected by western blot. (E) Flow cytometry was applied to measure cell apoptosis. Data represent the mean values ± standard deviation of three experiments. ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; OE-Wnt4, overexpression of Wnt4, OE-NC, negative control.

Techniques: Negative Control, Transfection, Western Blot, Immunofluorescence, Staining, Flow Cytometry, Standard Deviation, Over Expression

NF-κB participates in the effect of Wnt4 on LPS-induced HDPCs. The expressions of (A) inflammatory factors and (B) apoptosis-related proteins were detected by western blot. (C) Flow cytometry was applied to measure cell apoptosis. Data represent the mean values ± standard deviation of three experiments. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; OE-Wnt4, overexpression of Wnt4, OE-NC, negative control.

Journal: Experimental and Therapeutic Medicine

Article Title: Wnt4 prevents apoptosis and inflammation of dental pulp cells induced by LPS by inhibiting the IKK/NF‑κB pathway

doi: 10.3892/etm.2022.11774

Figure Lengend Snippet: NF-κB participates in the effect of Wnt4 on LPS-induced HDPCs. The expressions of (A) inflammatory factors and (B) apoptosis-related proteins were detected by western blot. (C) Flow cytometry was applied to measure cell apoptosis. Data represent the mean values ± standard deviation of three experiments. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. LPS, lipopolysaccharide; HDPCs, human dental pulp cells; OE-Wnt4, overexpression of Wnt4, OE-NC, negative control.

Techniques: Western Blot, Flow Cytometry, Standard Deviation, Over Expression, Negative Control

Schematic model for the molecular mechanism of Wnt4 in LPS-induced HDPCs. LPS, lipopolysaccharide; HDPCs, human dental pulp cells.

Journal: Experimental and Therapeutic Medicine

Article Title: Wnt4 prevents apoptosis and inflammation of dental pulp cells induced by LPS by inhibiting the IKK/NF‑κB pathway

doi: 10.3892/etm.2022.11774

Figure Lengend Snippet: Schematic model for the molecular mechanism of Wnt4 in LPS-induced HDPCs. LPS, lipopolysaccharide; HDPCs, human dental pulp cells.

Techniques:

Serum Wnt2 and Wnt4 are increased in patients with AMI and correlated to the increased risk of adverse outcomes of patients (a, b) ELISA analysis of serum Wnt2 and Wnt4 level in a total of 109 patients with AMI and 56 non-AMI patients. Data are expressed as means±SD, * p < 0.05, ** p < 0.01, *** p < 0.001 by Student's t test. (c, d) Kaplan–Meier incidence of MACEs in one year according to high or low level of serum Wnt2 or Wnt4. Wnt2 and Wnt4 were dichotomized into 2 categories with categorical analysis including higher than median and lower than median. Wnt2 high: ≥0.86 (ng/mL); Wnt2 low: < 0.86(ng/mL); Wnt4 high:≥86.2(pg/mL); Wnt4 low: < 86.2(pg/mL). MACEs: major adverse cardiovascular events. Estimated HR, 95% CIs, and p values were calculated. Statistics: The cumulative incidence of the MACE was determined by the Kaplan–Meier method, and the difference between groups was compared using the log-rank test. MACEs: major adverse cardiovascular events. Estimated HR, 95% CIs, and p values were calculated.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Serum Wnt2 and Wnt4 are increased in patients with AMI and correlated to the increased risk of adverse outcomes of patients (a, b) ELISA analysis of serum Wnt2 and Wnt4 level in a total of 109 patients with AMI and 56 non-AMI patients. Data are expressed as means±SD, * p < 0.05, ** p < 0.01, *** p < 0.001 by Student's t test. (c, d) Kaplan–Meier incidence of MACEs in one year according to high or low level of serum Wnt2 or Wnt4. Wnt2 and Wnt4 were dichotomized into 2 categories with categorical analysis including higher than median and lower than median. Wnt2 high: ≥0.86 (ng/mL); Wnt2 low: < 0.86(ng/mL); Wnt4 high:≥86.2(pg/mL); Wnt4 low: < 86.2(pg/mL). MACEs: major adverse cardiovascular events. Estimated HR, 95% CIs, and p values were calculated. Statistics: The cumulative incidence of the MACE was determined by the Kaplan–Meier method, and the difference between groups was compared using the log-rank test. MACEs: major adverse cardiovascular events. Estimated HR, 95% CIs, and p values were calculated.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Univariate and multivariable adjusted Cox proportional hazard regression models of incident MACEs with serum Wnt2 and Wnt4 in AMI patients.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Univariate and multivariable adjusted Cox proportional hazard regression models of incident MACEs with serum Wnt2 and Wnt4 in AMI patients.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques:

Serum and cardiac Wnt2 and Wnt4 levels are increased in mice following MI. (a) Western blot analysis of the expression of Wnt2, Wnt4, Col1, Col3 and TGF- β1 in the border zone of infarcted area at different time-points (3d, 7d, 14d, 28d) following MI. Sham operation was used as control. Values were expressed as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001 vs sham group, n = 5/group. (b) Western blot analysis of serum Wnt2 and Wnt4 from sham group and different time-points (3d,7d,14d,28d) after MI. Values were expressed as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001 vs sham group, n = 4/group. (c) Wnt2 and Wnt4 expression were analyzed by Western blot analysis in cultured neonatal rat cardiac fibroblasts (NRCFs) at different time-points (1 h,3 h,6 h,12 h,24 h) in response to hypoxia. Normaxia group was used as control. Values were expressed as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001 vs control group. n = 4/group. (d) The representative images of Wnt2 and Wnt4 expression with Western blot analysis in conditional medium from neonatal rat cardiac fibroblasts (NRCFs) of control group and at different time-points (1 h,3 h,6 h,12 h,24 h) after hypoxia. In (b) and (d), a conventional Coomassie staining of polyvinylidene fluoride (PVDF) membranes showed the total protein load which was as the internal control (Loading control). The experiment was repeated for three times. MI: Myocardial infarction; CON: control. Statistics: One-way ANOVA with post-hoc Tukey test.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Serum and cardiac Wnt2 and Wnt4 levels are increased in mice following MI. (a) Western blot analysis of the expression of Wnt2, Wnt4, Col1, Col3 and TGF- β1 in the border zone of infarcted area at different time-points (3d, 7d, 14d, 28d) following MI. Sham operation was used as control. Values were expressed as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001 vs sham group, n = 5/group. (b) Western blot analysis of serum Wnt2 and Wnt4 from sham group and different time-points (3d,7d,14d,28d) after MI. Values were expressed as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001 vs sham group, n = 4/group. (c) Wnt2 and Wnt4 expression were analyzed by Western blot analysis in cultured neonatal rat cardiac fibroblasts (NRCFs) at different time-points (1 h,3 h,6 h,12 h,24 h) in response to hypoxia. Normaxia group was used as control. Values were expressed as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001 vs control group. n = 4/group. (d) The representative images of Wnt2 and Wnt4 expression with Western blot analysis in conditional medium from neonatal rat cardiac fibroblasts (NRCFs) of control group and at different time-points (1 h,3 h,6 h,12 h,24 h) after hypoxia. In (b) and (d), a conventional Coomassie staining of polyvinylidene fluoride (PVDF) membranes showed the total protein load which was as the internal control (Loading control). The experiment was repeated for three times. MI: Myocardial infarction; CON: control. Statistics: One-way ANOVA with post-hoc Tukey test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Western Blot, Expressing, Control, Cell Culture, Staining

$Knockdown of Wnt2/Wnt4 suppresses cardiac dysfunction and fibrosis post-MI. 8-10 weeks old male mice were injected with shWnt2/4-AAV9 (shWnt2/4) or shScramble-AAV9 (shNC, as control) by tail vein at 3 weeks before sham or MI operation. (a) Wnt2 and Wnt4 levels were determined in the non-infarcted area by western blot method on day 28 post-MI. Values were expressed as means±S.E.M; *** p < 0.001 vs shNC group; n = 5/each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. (b) Echocardiographic analysis of mice at day 0, day7, day14 and day28 after MI. Left lane: Representative images of M-mode echocardiogram captured on the 28th day post-MI. Right lane: quantitative analysis of LVEF, FS; LVEF: Left ventricular ejection fraction; FS, fraction shortening. Values were presented as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001 vs shNC group; n = 6/group. Statistics: Student's t test. (c) Hemodynamics analysis of dp/dt, -dp/dt, LVEDP and Tau at 21 days after MI in mice. Values were expressed as means±S.E.M. * p < 0.05;** p < 0.01;*** p < 0.001, n = 4-8 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. (d) Cardiac fibrosis was examined by Masson staining at 28 days after MI in mice. Bar =1000 μm. Values were expressed as means±S.E.M. ** p < 0.01 vs shNC group, n = 6 in each group. Statistics: Student's t test. (e) Col1, Col3, MMP2, MMP9 and α-SMA protein levels were analyzed by Western blot in border zone of infarcted area on day 28 post-MI. Values were presented as means±S.E.M .* p < 0.05, ** p < 0.01,*** p < 0.001; n = 4 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.$

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Knockdown of Wnt2/Wnt4 suppresses cardiac dysfunction and fibrosis post-MI. 8-10 weeks old male mice were injected with shWnt2/4-AAV9 (shWnt2/4) or shScramble-AAV9 (shNC, as control) by tail vein at 3 weeks before sham or MI operation. (a) Wnt2 and Wnt4 levels were determined in the non-infarcted area by western blot method on day 28 post-MI. Values were expressed as means±S.E.M; *** p < 0.001 vs shNC group; n = 5/each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. (b) Echocardiographic analysis of mice at day 0, day7, day14 and day28 after MI. Left lane: Representative images of M-mode echocardiogram captured on the 28th day post-MI. Right lane: quantitative analysis of LVEF, FS; LVEF: Left ventricular ejection fraction; FS, fraction shortening. Values were presented as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001 vs shNC group; n = 6/group. Statistics: Student's t test. (c) Hemodynamics analysis of dp/dt, -dp/dt, LVEDP and Tau at 21 days after MI in mice. Values were expressed as means±S.E.M. * p < 0.05;** p < 0.01;*** p < 0.001, n = 4-8 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. (d) Cardiac fibrosis was examined by Masson staining at 28 days after MI in mice. Bar =1000 μm. Values were expressed as means±S.E.M. ** p < 0.01 vs shNC group, n = 6 in each group. Statistics: Student's t test. (e) Col1, Col3, MMP2, MMP9 and α-SMA protein levels were analyzed by Western blot in border zone of infarcted area on day 28 post-MI. Values were presented as means±S.E.M .* p < 0.05, ** p < 0.01,*** p < 0.001; n = 4 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Knockdown, Injection, Control, Western Blot, Staining

Wnt2 or Wnt4 is involved in fibroblasts activation in response to hypoxia. (a,b) Western blot analysis of the expressions of Wnt2, Wnt4, Col1, Col3, TGFβ1, MMP9 MMP2, p-smad2/3, CTGF and α-SMA protein in neonatal rat cardiac fibroblasts (NRCFs). These NRCFs were pretreated with siRNA targeted Wnt2 or Wnt4 (si-Wnt2 or si-Wnt4) or si-Scramble (si-Scram) for 24 h, and then exposed to hypoxia or normoxia (Control) for 3 h. p: pro-TGF-β1; m: mature or active TGF-β1; TGF-β1 mature form was analyzed. Values were described as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001; n = 3/group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. c: Scratch assay for migratory ability of cardiac fibroblasts pre-transfected with si-scram (CON), si-Wnt2 or si-Wnt4 followed by hypoxia. The representative images were showed at 0 h and 12 h after hypoxia. The bar=20 um; Values were quantitified and expressed as means±S.E.M. ** p < 0.01, *** p < 0.001 vs si-scram group; n = 4/group. Statistics: One-way ANOVA with post-hoc Tukey test.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Wnt2 or Wnt4 is involved in fibroblasts activation in response to hypoxia. (a,b) Western blot analysis of the expressions of Wnt2, Wnt4, Col1, Col3, TGFβ1, MMP9 MMP2, p-smad2/3, CTGF and α-SMA protein in neonatal rat cardiac fibroblasts (NRCFs). These NRCFs were pretreated with siRNA targeted Wnt2 or Wnt4 (si-Wnt2 or si-Wnt4) or si-Scramble (si-Scram) for 24 h, and then exposed to hypoxia or normoxia (Control) for 3 h. p: pro-TGF-β1; m: mature or active TGF-β1; TGF-β1 mature form was analyzed. Values were described as means±S.E.M; * p < 0.05, ** p < 0.01, *** p < 0.001; n = 3/group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. c: Scratch assay for migratory ability of cardiac fibroblasts pre-transfected with si-scram (CON), si-Wnt2 or si-Wnt4 followed by hypoxia. The representative images were showed at 0 h and 12 h after hypoxia. The bar=20 um; Values were quantitified and expressed as means±S.E.M. ** p < 0.01, *** p < 0.001 vs si-scram group; n = 4/group. Statistics: One-way ANOVA with post-hoc Tukey test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Activation Assay, Western Blot, Control, Wound Healing Assay, Transfection

Wnt2 or Wnt4 activates NF-κB pathway signaling and enhances the expression of Fzd4/Fzd2 in neonatal rat cardiac fibroblasts (NRCFs) in response to hypoxia. (a) The expressions of p-p65, P65, Fzd2 and Fzd4 in NRCFs were detected by Western blot analysis. n = 4/group. (b) The expressions of p65 and β-catenin were detected in nucleus of NRCFs by western blot. n = 3/group. (a,b) These NRCFs were pretreated with siRNA targeted Wnt2 or Wnt4 (si-Wnt2 or si-Wnt4) or si-Scramble (si-Scram) for 24 h, and then exposed to hypoxia or normoxia (Control) for 3 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001. (c) The expressions of p-p65, p65, Fzd 2 and Fzd 4 were detected in non-infarcted area by Western blot analysis. 8-10 weeks old male mice were intravenously injected with shWnt2/4-AAV9 (shWnt2/4) or shScramble-AAV9 (shNC) at 3 weeks before sham or MI operation. These proteins were detected at day 28 post MI or sham operation. n = 4 in each group. Values were expressed as means±S.E.M; ** p < 0.01, *** p < 0.001. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Wnt2 or Wnt4 activates NF-κB pathway signaling and enhances the expression of Fzd4/Fzd2 in neonatal rat cardiac fibroblasts (NRCFs) in response to hypoxia. (a) The expressions of p-p65, P65, Fzd2 and Fzd4 in NRCFs were detected by Western blot analysis. n = 4/group. (b) The expressions of p65 and β-catenin were detected in nucleus of NRCFs by western blot. n = 3/group. (a,b) These NRCFs were pretreated with siRNA targeted Wnt2 or Wnt4 (si-Wnt2 or si-Wnt4) or si-Scramble (si-Scram) for 24 h, and then exposed to hypoxia or normoxia (Control) for 3 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001. (c) The expressions of p-p65, p65, Fzd 2 and Fzd 4 were detected in non-infarcted area by Western blot analysis. 8-10 weeks old male mice were intravenously injected with shWnt2/4-AAV9 (shWnt2/4) or shScramble-AAV9 (shNC) at 3 weeks before sham or MI operation. These proteins were detected at day 28 post MI or sham operation. n = 4 in each group. Values were expressed as means±S.E.M; ** p < 0.01, *** p < 0.001. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Expressing, Western Blot, Control, Injection

NF-κB inhibitor ameliorates Wnt2/Wnt4-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) The expressions of Col1, Col3, active β-catenin, MMP2, MMP9, p65, p-p65 and TGFβ1 were measured in NRCFs by Western blot analysis. These NRCFs were pretreated with JSH-23(NF-κB inhibitor, 10μM) or the same volume of DMSO for 1 h, followed by PBS, human recombinant Wnt2 (20 ng/ml) or Wnt4 (50 ng/ml) treatment for 24 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001; n = 3 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: NF-κB inhibitor ameliorates Wnt2/Wnt4-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) The expressions of Col1, Col3, active β-catenin, MMP2, MMP9, p65, p-p65 and TGFβ1 were measured in NRCFs by Western blot analysis. These NRCFs were pretreated with JSH-23(NF-κB inhibitor, 10μM) or the same volume of DMSO for 1 h, followed by PBS, human recombinant Wnt2 (20 ng/ml) or Wnt4 (50 ng/ml) treatment for 24 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001; n = 3 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Western Blot, Recombinant

Wnt/β-catenin inhibitor relieves Wnt2/Wnt4-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) The expressions of Col1, Col3, active β-catenin, p65, TGFβ1, Fzd2, Fzd4 and p-p65 were measured in NRCFs by Western blot analysis. These NRCFs were pretreated with ICG-001(β-catenin inhibitor, 10 μM) or the same volume of DMSO for 1 h, followed by PBS, human recombinant Wnt2 (20 ng/ml) or Wnt4 (50 ng/ml) treatment for 24 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 3 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Wnt/β-catenin inhibitor relieves Wnt2/Wnt4-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) The expressions of Col1, Col3, active β-catenin, p65, TGFβ1, Fzd2, Fzd4 and p-p65 were measured in NRCFs by Western blot analysis. These NRCFs were pretreated with ICG-001(β-catenin inhibitor, 10 μM) or the same volume of DMSO for 1 h, followed by PBS, human recombinant Wnt2 (20 ng/ml) or Wnt4 (50 ng/ml) treatment for 24 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 3 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Western Blot, Recombinant

Knockdown of Fzd2 or Fzd4 attenuates Wnt4- or Wnt2-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) Western blot analysis of the expressions of Fzd2, Col1, Col3, active β-catenin, MMP2, MMP9, TGFβ1, and p-65 in NRCFs. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001; n = 3 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. (a) NRCFs were pretreated with siRNA targeted Fzd2 (si- Fzd2) or si-Scramble (si-Scram) for 24 h, followed by PBS, human recombinant Wnt4 (50 ng/ml) treatment for another 24 h. (b) NRCFs were pretreated with siRNA targeted Fzd4 (si- Fzd4) or si-Scramble (si-Scram) for 24 h, followed by PBS, human recombinant Wnt2 (20 ng/ml) treatment for another 24 h.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Knockdown of Fzd2 or Fzd4 attenuates Wnt4- or Wnt2-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) Western blot analysis of the expressions of Fzd2, Col1, Col3, active β-catenin, MMP2, MMP9, TGFβ1, and p-65 in NRCFs. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001; n = 3 in each group. Statistics: Two-way ANOVA with a Bonferroni post hoc test. (a) NRCFs were pretreated with siRNA targeted Fzd2 (si- Fzd2) or si-Scramble (si-Scram) for 24 h, followed by PBS, human recombinant Wnt4 (50 ng/ml) treatment for another 24 h. (b) NRCFs were pretreated with siRNA targeted Fzd4 (si- Fzd4) or si-Scramble (si-Scram) for 24 h, followed by PBS, human recombinant Wnt2 (20 ng/ml) treatment for another 24 h.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Knockdown, Western Blot, Recombinant

Knockdown of LRP6 attenuates Wnt2- or Wnt4-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) Western blot analysis of the expressions of Col1, Col3, active β-catenin, TGFβ1, LRP6 and p-65 in NRCFs. Cultured NRCFs were transfected with lenti-shScr (shScr) or lenti-LRP6(shLRP6) for 48 h and then followed by PBS, human recombinant Wnt2 (20 ng/ml) or Wnt4(50 ng/ml) treatment for another 24 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001. n = 3 /group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: Knockdown of LRP6 attenuates Wnt2- or Wnt4-induced pro-fibrotic effects in neonatal rat cardiac fibroblasts (NRCFs). (a,b) Western blot analysis of the expressions of Col1, Col3, active β-catenin, TGFβ1, LRP6 and p-65 in NRCFs. Cultured NRCFs were transfected with lenti-shScr (shScr) or lenti-LRP6(shLRP6) for 48 h and then followed by PBS, human recombinant Wnt2 (20 ng/ml) or Wnt4(50 ng/ml) treatment for another 24 h. Values were expressed as means±S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001. n = 3 /group. Statistics: Two-way ANOVA with a Bonferroni post hoc test.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Knockdown, Western Blot, Cell Culture, Transfection, Recombinant

A proposed model of showing how Wnt2 and Wnt4 promote cardiac fibrosis by activation of β-catenin/NF-κB trough LRP6 and Fzds signaling following MI. MI induced the increased expression or secretion of Wnt2 and Wnt4 in cardiaomyocytes (CMs) or cardiac fibroblasts (CFs), elevated Wnt2 and Wnt4 promote fibrotic effects by activation of β-catenin/NF-κB signaling dependently on the cooperation of Fzd4 or Fzd2 and LRP6 signaling in cardiac fibroblasts, which contributes to cardiac fibrosis and dysfunction post-MI. Thus high Wnt2 and Wnt4 are independently associated with adverse outcome in AMI patients.

Journal: EBioMedicine

Article Title: Elevated Wnt2 and Wnt4 activate NF-κB signaling to promote cardiac fibrosis by cooperation of Fzd4/2 and LRP6 following myocardial infarction

doi: 10.1016/j.ebiom.2021.103745

Figure Lengend Snippet: A proposed model of showing how Wnt2 and Wnt4 promote cardiac fibrosis by activation of β-catenin/NF-κB trough LRP6 and Fzds signaling following MI. MI induced the increased expression or secretion of Wnt2 and Wnt4 in cardiaomyocytes (CMs) or cardiac fibroblasts (CFs), elevated Wnt2 and Wnt4 promote fibrotic effects by activation of β-catenin/NF-κB signaling dependently on the cooperation of Fzd4 or Fzd2 and LRP6 signaling in cardiac fibroblasts, which contributes to cardiac fibrosis and dysfunction post-MI. Thus high Wnt2 and Wnt4 are independently associated with adverse outcome in AMI patients.

Article Snippet: Human Wnt2 and Wnt4 ELISA kits were purchased from CUSABIO company (Cat#: CSB-EL026133HU, CSB-EL026137HU, Wuhan, China) and used following the manufacturer's instructions.

Techniques: Activation Assay, Expressing

Journal: Jcsm Rapid Communications

Article Title: Skeletal muscle‐specific over‐expression of the nuclear sirtuin SIRT6 blocks cancer‐associated cachexia by regulating multiple targets

doi: 10.1002/rco2.27

Figure Lengend Snippet: Primer sequences used in this study

Article Snippet: WNT4 concentration in plasma was estimated using 10–20 μL plasma and mouse protein WNT4 ELISA kit (MyBioSource Inc., MBS903402) as per manufacturer's instructions.

Techniques: Sequencing

Journal: Jcsm Rapid Communications

Article Title: Skeletal muscle‐specific over‐expression of the nuclear sirtuin SIRT6 blocks cancer‐associated cachexia by regulating multiple targets

doi: 10.1002/rco2.27

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: WNT4 concentration in plasma was estimated using 10–20 μL plasma and mouse protein WNT4 ELISA kit (MyBioSource Inc., MBS903402) as per manufacturer's instructions.

Techniques:

Over‐expressed SIRT6 also targets WNT4 in gastrocnemius muscle. ( A ) no significant change in Wnt4 mRNA expression in non‐tumour background but ( B ) SIRT6 downregulates Wnt4 (normalized to RNA polymerase 2 ) mRNA expression in tumour setting ( n = 5–7 mice). Representative western blots showing WNT4 protein expression for both mice categories in non‐tumour ( C ), and ( D ) tumour‐bearing conditions ( n = 5–8). Coomassie blue‐stained blots are shown as loading controls. Bar graphs showing plasma WNT4 concentrations for non‐tumour ( E ) ( n = 7–10), and tumour‐bearing ( F ) control and Tg mice ( n = 10–20 mice). Bar graphs represent data as mean ± SEM. NS, non‐significant. * P < 0.05.

Journal: Jcsm Rapid Communications

Article Title: Skeletal muscle‐specific over‐expression of the nuclear sirtuin SIRT6 blocks cancer‐associated cachexia by regulating multiple targets

doi: 10.1002/rco2.27

Figure Lengend Snippet: Over‐expressed SIRT6 also targets WNT4 in gastrocnemius muscle. ( A ) no significant change in Wnt4 mRNA expression in non‐tumour background but ( B ) SIRT6 downregulates Wnt4 (normalized to RNA polymerase 2 ) mRNA expression in tumour setting ( n = 5–7 mice). Representative western blots showing WNT4 protein expression for both mice categories in non‐tumour ( C ), and ( D ) tumour‐bearing conditions ( n = 5–8). Coomassie blue‐stained blots are shown as loading controls. Bar graphs showing plasma WNT4 concentrations for non‐tumour ( E ) ( n = 7–10), and tumour‐bearing ( F ) control and Tg mice ( n = 10–20 mice). Bar graphs represent data as mean ± SEM. NS, non‐significant. * P < 0.05.

Article Snippet: WNT4 concentration in plasma was estimated using 10–20 μL plasma and mouse protein WNT4 ELISA kit (MyBioSource Inc., MBS903402) as per manufacturer's instructions.

Techniques: Expressing, Western Blot, Staining, Clinical Proteomics, Control

WNT4 promote angiogenesis in colorectal cancer. a and b The expression of CD31 (red) in different groups: LoVo-Scramble or LoVo-shWNT4 ( a ), SW480-WNT4-vector or SW480-WNT4-HA. Scale bar, 50 μm. c The ability of tube formation was significantly improved when HUVECs were treated with conditioned media (CM) from SW480 cells transfected with WNT4-HA than that from SW480 transfected WNT4-vector, and it could be blocked by the β-catenin/TCF inhibitor ICG-001 (10 μM). Scale bar, 100 μm. d TOP/FOP-Flash reporter was used to detect whether the WNT pathway was activated. e and f The levels of ANG2 were detected by RT-qPCR and ELISA in HUVECs that underwent different treatments. g and h The mRNA level of ANG2 in cells and CM of HUVECs transfected with negative control shRNA (Scramble) and sh-ANG2 were detected by RT-qPCR. i The ability of tube formation promoted by high WNT4 expression was significantly inhibited due to low ANG2 expression. Scale bar, 100 μm. ** P < 0.01

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: WNT4 secreted by tumor tissues promotes tumor progression in colorectal cancer by activation of the Wnt/β-catenin signalling pathway

doi: 10.1186/s13046-020-01774-w

Figure Lengend Snippet: WNT4 promote angiogenesis in colorectal cancer. a and b The expression of CD31 (red) in different groups: LoVo-Scramble or LoVo-shWNT4 ( a ), SW480-WNT4-vector or SW480-WNT4-HA. Scale bar, 50 μm. c The ability of tube formation was significantly improved when HUVECs were treated with conditioned media (CM) from SW480 cells transfected with WNT4-HA than that from SW480 transfected WNT4-vector, and it could be blocked by the β-catenin/TCF inhibitor ICG-001 (10 μM). Scale bar, 100 μm. d TOP/FOP-Flash reporter was used to detect whether the WNT pathway was activated. e and f The levels of ANG2 were detected by RT-qPCR and ELISA in HUVECs that underwent different treatments. g and h The mRNA level of ANG2 in cells and CM of HUVECs transfected with negative control shRNA (Scramble) and sh-ANG2 were detected by RT-qPCR. i The ability of tube formation promoted by high WNT4 expression was significantly inhibited due to low ANG2 expression. Scale bar, 100 μm. ** P < 0.01

Article Snippet: The WNT4 concentration in serum from healthy donors, CRC patients, and conditioned medium from fresh tissues were quantitatively determined by ELISA kits (CSB-EL026137HU, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Expressing, Plasmid Preparation, Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Negative Control, shRNA

Loss of Gpr177 blocked the extracellular secretion of WNT proteins. ( a– c ) Protein levels of WNT3 ( a ), WNT3A ( b ) and WNT7A ( c ) in germ cell culture supernatants from control, Gpr177 flox/flox , Mvh -Cre; Gpr177 flox/flox , Stra8 -Cre and Gpr177 flox/flox , Amh -Cre testes. ( d – f ) Secreted WNT4 ( d ), WNT6 ( e ) and WNT11 ( f ) examined by ELISA methods were observed in supernatants of Sertoli cells from control and three Gpr177 cKO testes. The data are expressed as the mean±S.E.M. ** P <0.01

Journal: Cell Death & Disease

Article Title: Does murine spermatogenesis require WNT signalling? A lesson from Gpr177 conditional knockout mouse models

doi: 10.1038/cddis.2016.191

Figure Lengend Snippet: Loss of Gpr177 blocked the extracellular secretion of WNT proteins. ( a– c ) Protein levels of WNT3 ( a ), WNT3A ( b ) and WNT7A ( c ) in germ cell culture supernatants from control, Gpr177 flox/flox , Mvh -Cre; Gpr177 flox/flox , Stra8 -Cre and Gpr177 flox/flox , Amh -Cre testes. ( d – f ) Secreted WNT4 ( d ), WNT6 ( e ) and WNT11 ( f ) examined by ELISA methods were observed in supernatants of Sertoli cells from control and three Gpr177 cKO testes. The data are expressed as the mean±S.E.M. ** P <0.01

Article Snippet: Supernatants were collected and used for ELISA analysis for quantification of secreted WNT3 (CSB-EL026135MO), WNT3A (CSB-EL026136MO), WNT7A (CSB-EL026141MO), WNT4 (CSB-EL026137MO), WNT6 (CSB-EL026140MO) and WNT11 (CSB-EL026131MO) (CUSABIO, Wuhan, China) using the manufacturer's instructions.

Techniques: Cell Culture, Control, Enzyme-linked Immunosorbent Assay

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