human wnt2 protein  (Novus Biologicals)

Journal: Advanced Science

Article Title: Optogenetic Stimulation of mPFC Alleviates White Matter Injury‐Related Cognitive Decline after Chronic Ischemia through Adaptive Myelination

doi: 10.1002/advs.202202976

Figure Lengend Snippet: Activation of glutamatergic neurons in the mPFC upregulates Wnt2 expression in the corpus callosum. a) Heatmap of the results of mPFC tissue microarray analysis between the BCAS mCherry and BCAS ChR2 groups at 2 months after surgery. n = 3 per group. b) Volcano plot of the results of mPFC tissue microarray analysis between the BCAS mCherry and BCAS ChR2 groups at 2 months after surgery (fold change > 2; p < 0.05). The DEGs are listed. n = 3 per group. c) Real‐time PCR for validation of the identified DEGs among the sham mCherry, BCAS mCherry, and BCAS ChR2 groups at 2 months after surgery. n = 7–8 per group. d) Real‐time PCR analysis of Wnt2 expression in the mPFC in the sham mCherry, BCAS mCherry, and BCAS hM3D groups at 2 months after surgery. n = 7–8 per group. e) Real‐time PCR analysis of Wnt2 expression in different types of primary cortical neural cells. n = 3 per group. f) Representative immunostaining of NeuN and Wnt2 in the mPFC in the sham mCherry, BCAS mCherry, and BCAS ChR2 groups at 2 months after surgery. Scale bar: 50 µm. g) Representative immunostaining of NF‐H and Wnt2 in the corpus callosum in the sham mCherry, BCAS mCherry, and BCAS ChR2 groups at 2 months after surgery. Scale bar: 35 µm. Quantification of immunostaining of Wnt2 in the h) mPFC and i) corpus callosum in the sham mCherry, BCAS mCherry, and BCAS ChR2 groups at 2 months after surgery. The fluorescence intensity in each image was normalized to that of NeuN and NF‐H. The measured values were normalized to the mean value of the BCAS group. n = 3 per group. j) Representative immunostaining of NeuN and Wnt2 in the mPFC in the sham mCherry, BCAS mCherry, and BCAS hM3D groups at 2 months after surgery. Scale bar: 50 µm. k) Representative immunostaining of NF‐H and Wnt2 in the corpus callosum in the sham mCherry, BCAS mCherry, and BCAS hM3D groups at 2 months after surgery. Scale bar: 35 µm. Quantification of Wnt2 immunostaining in the l) mPFC and m) corpus callosum in the sham mCherry, BCAS mCherry, and BCAS hM3D groups at 2 months after surgery. The fluorescence intensity in each image was normalized to that of NeuN and NF‐H. The measured values were normalized to the mean value of the BCAS group. n = 3 per group. The data are presented as the mean ± SEM. p ‐values were determined by the Kruskal–Wallis test with Dunn's post‐hoc analysis in (d); by 1‐way ANOVA with Tukey's post‐hoc analysis in (c) and (e); and by Student's t ‐test in (h), (i), (l), and (m). * p < 0.05, ** p < 0.01.

Article Snippet: Afterward, 50 ng mL −1 T3 (Thermo Fisher Scientific, USA), 200 ng mL −1 recombinant Wnt2 protein (Cusabio, China), 100 ng mL −1 recombinant Dkk1 protein (Cusabio, China) and basic medium DMEM F12 supplemented with 2% B27, 100 U mL −1 antibiotics, and 20 ng mL −1 CNTF (Thermo Fisher Scientific, USA) were applied to induce differentiation.

Techniques: Activation Assay, Expressing, Microarray, Real-time Polymerase Chain Reaction, Biomarker Discovery, Immunostaining, Fluorescence

Journal: Advanced Science

Article Title: Optogenetic Stimulation of mPFC Alleviates White Matter Injury‐Related Cognitive Decline after Chronic Ischemia through Adaptive Myelination

doi: 10.1002/advs.202202976

Figure Lengend Snippet: Overexpression of Wnt2 in mPFC glutamatergic neurons alleviates myelin injury and improves cognition. Results of the a) Y‐maze and b) T‐maze tests showing the spontaneous alternation percentage in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 15–16 per group. Results of the open field test showing c) the total distance travelled and d) time spent in the corner area and e) center area in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 15–16 per group. f) Heatmaps generated from DTI axial views of FA acquired from the control and Camk2a‐Wnt2 groups at 2 months after surgery. g) Quantification of FA values in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. The measured values were normalized to the mean value of the control group. n = 11 per group. h) Representative images of Black‐gold staining and immunostaining of MBP and MAG in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Scale bar: 200 µm. Quantification of i) Black‐gold staining and immunostaining of j) MBP and k) MAG in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 4 per group. l) Representative TEM images of the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Scale bar: 1 µm. m) The percentage of myelinated axons in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 4 per group. n) Scatterplots of the myelin g‐ratio as a function of the axon diameter in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Axons were selected from 4 mice per group for measurement. n = 138 and n = 121 measured axons for each group, respectively. o) Representative immunostaining of CC1 and Olig2 in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. Scale bar: 50 µm. Density of p) CC1 + Olig2 + cells and q) CC1 − Olig2 + cells in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 5 per group. r) Proportion of CC1 + /Olig2 + cells in the corpus callosum in the control and Camk2a‐Wnt2 groups at 2 months after surgery. n = 5 per group. s) Immunoblot bands and t) quantification of MBP and u) MAG expression in OPCs in the control, T3, and T3+ rmWnt2 groups. The intensity of each immunoblot band was normalized to that of the β ‐actin band. The measured values were normalized to the mean value of the control group. n = 3 per group. The data are presented as the mean ± SEM. p ‐values were determined by Student's t ‐test in (b–e), (i–k), (m), (p–r), and (t); by the Mann–Whitney test in (a), (g), and (n); and by 1‐way ANOVA with Tukey's post‐hoc analysis in (u). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Afterward, 50 ng mL −1 T3 (Thermo Fisher Scientific, USA), 200 ng mL −1 recombinant Wnt2 protein (Cusabio, China), 100 ng mL −1 recombinant Dkk1 protein (Cusabio, China) and basic medium DMEM F12 supplemented with 2% B27, 100 U mL −1 antibiotics, and 20 ng mL −1 CNTF (Thermo Fisher Scientific, USA) were applied to induce differentiation.

Techniques: Over Expression, Control, Generated, Staining, Immunostaining, Western Blot, Expressing, MANN-WHITNEY

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

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:

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

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

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

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

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

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

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

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

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: 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

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:

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

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

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

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

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

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

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

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

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: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: Primer sequences for RT-qPCR

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Sequencing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: The potential involvement of the LINC00968/HEY1/WNT2 axis mediated Wnt2/β-catenin signaling pathway in breast cancer. a selection thermogram of GEO dataset GSE26910. The horizontal coordinate represents the samples, and the vertical coordinate represents the genes. The upper right histogram is the color gradation. Each rectangle corresponds to one gene expression in one sample. Each column signifies gene expression of each sample; dendrogram on the left side represents cluster analysis of different genes from different samples; uppermost crossband represents sample type and upper right pane signifies color reference, while blue is the normal control sample and red is tumor sample. b the expression pattern of LINC00968 in cancer tissues and normal tissues of various cancers according to TCGA database. c the expression of LINC00968 in breast cancer tissues and adjacent normal tissues. d the survival analysis of LINC00968 expression and survival rate of patients. e the subcellular localization of LINC00968 in breast cancer cells as detected by FISH assay. f the binding status of LINC00968 to HEY1 as confirmed by RIP assay. g the binding sites of HEY1 to the WNT2 promoter as predicted by online Jaspar website ( http://jaspar.genereg.net/ ). h the binding status of HEY1 to the WNT2 promoter as verified by ChIP assay. i the interaction between LINC00968 and WNT2 as verified by Dual luciferase reporter gene assay. * p < 0.05 vs. the NC group. j the mRNA expression of INC00968 and WNT2 as determined by RT-qPCR. k the protein levels of INC00968 and WNT2 as evaluated by Western blot analysis. l the expression of WNT2 in cell culture medium as assayed by ELISA. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector group. m the mRNA expression of LINC00968, WNT2, and β-catenin in breast cancer tissues and normal adjacent tissues as measured by RT-qPCR. n the protein level of WNT2 and the phosphorylation levels of β-catenin and GSK3β. o the level of LINC00968 determined by Northern blot analysis. * p < 0.05 vs. the adjacent normal tissues

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Selection, Gene Expression, Control, Expressing, Binding Assay, Luciferase, Reporter Gene Assay, Quantitative RT-PCR, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Phospho-proteomics, Northern Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: KEGG pathway enrichment analysis of the target genes of LINC00968

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: LINC00968 contributes to inhibition of the Wnt2/β-catenin signaling pathway, elevated E-cadherin level and downregulated Vimentin level. a the mRNA expression of β-catenin, GSK3β and Vimentin in MCF-7/ADM cells after plasmid delivery in each group as measured by RT-qPCR. b protein bands of β-catenin/p-β-catenin, GSK3β/p-GSK3β and Vimentin/p-Vimentin in MCF-7/ADM cells after plasmid delivery of each group as determined by Western blot analysis. c histogram of levels of β-catenin/p-β-catenin, GSK3β/p-GSK3β and Vimentin/p-Vimentin in each group. d protein bands of LINC00968 in MCF-7/ADM cells after plasmid delivery of each group. e LINC00968 protein level in each group detected by Northern blot analysis. f the mRNA expression of β-catenin, GSK3β and Vimentin in KPL-4/ADM cells after plasmid delivery in each group as measured by RT-qPCR. g protein bands of β-catenin/p-β-catenin, GSK3β/p-GSK3β and Vimentin/p-Vimentin in KPL-4/ADM cells after plasmid delivery of each group as determined by Western blot analysis. h histogram of levels of β-catenin/p-β-catenin, GSK3β/p-GSK3β and Vimentin/p-Vimentin in each group. i protein bands of LINC00968 in KPL-4/ADM cells after plasmid delivery of each group as evaluated by Northern blot analysis. j histogram of LINC00968 protein level in each group. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector + WNT2 vector group

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Inhibition, Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Northern Blot, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: Overexpressed LINC00968 or silenced WNT2 contributes to reduced drug resistance of breast cancer cells. a drug resistance of transfected MCF-7/ADM cells to ADR as evaluated by CCK-8 assay. b drug resistance of transfected MCF-7/ADM cells to Taxel as determined by CCK-8 assay. c drug resistance of transfected MCF-7/ADM cells to VCR as assessed by CCK-8 assay. d protein bands of MRP1, BCRP, and P-gp in transfected MCF-7/ADM cells ad determined by Western blot analysis. e histogram of MRP1, BCRP, and P-gp protein levels in each group. f drug resistance of transfected KPL-4/ADM cells to ADR as evaluated by CCK-8 assay. g drug resistance of transfected KPL-4/ADM cells to Taxel as determined by CCK-8 assay. h drug resistance of transfected KPL-4/ADM cells to VCR as assessed by CCK-8 assay. i protein bands of MRP1, BCRP, and P-gp in transfected KPL-4/ADM cells ad determined by Western blot analysis. j histogram of MRP1, BCRP, and P-gp protein levels in each group. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector + WNT2 vector group

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Transfection, CCK-8 Assay, Western Blot, Control, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: Overexpressed LINC00968 or silenced WNT2 decrease colony formation ability of breast cancer cells. a the colony formation ability of transfected MCF-7 cells in each group as assessed by clonogenic assay. b histogram of the number of forming colony units in each group. c the colony formation ability of transfected KPL-4 cells in each group as evaluated by clonogenic assay. d histogram of the number of forming colony units in each group. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector + WNT2 vector group

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Transfection, Clonogenic Assay, Control, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: Overexpressed LINC00968 or silenced WNT2 decreases migration and invasion abilities of breast cancer cells. a the migration ability of MCF-7 cells after plasmid delivery in each group as evaluated by the scratch test. b the invasion ability of MCF-7 and MCF-7/ADM cells after plasmid delivery in each group as determined by the Transwell assay. c the cell migration rate in each group. d the number of invasive cells in each group. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector + WNT2 vector group

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Migration, Plasmid Preparation, Transwell Assay, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: Overexpressed LINC00968 or silenced WNT2 contributes to promoted apoptosis of breast cancer cells. a the apoptosis of MCF-7 cells after transduction in each group as analyzed by flow cytometry. b the cell apoptosis rate in each group. c protein bands of Bax, cleaved-PARP, cleaved-caspase3 and Bcl-2 in MCF-7/ADM cells after transduction in each group as measured by Western blot analysis. d the protein levels of Bax, cleaved-PARP, cleaved-caspase3 and Bcl-2 in each group. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector + WNT2 vector group

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Transduction, Flow Cytometry, Western Blot, Control, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: Overexpressed LINC00968 or silenced WNT2 restrains transplantation tumor growth in nude mice. a the tumor volume in nude mice of each group b the tumor weight in nude mice of each group. * p < 0.05 vs. the control group; # p < 0.05 vs. the blank and NC groups; & p < 0.05 vs. the LINC00968 vector + WNT2 vector group

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Transplantation Assay, Control, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: LINC00968 was lowly expressed, whereas WNT2 was highly expressed in breast cancer tissues and cells. LINC00968 targeted and negatively regulated WNT2 via the transcriptional repressor HEY1. Overexpressed LINC00968 reduced drug resistance, migration, invasion and epithelial-mesenchymal transition of breast cancer cells through inhibiting the activation of the Wnt2/β-catenin signaling pathway via suppression of WNT2

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Migration, Activation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long non-coding RNA LINC00968 attenuates drug resistance of breast cancer cells through inhibiting the Wnt2/β-catenin signaling pathway by regulating WNT2

doi: 10.1186/s13046-019-1100-8

Figure Lengend Snippet: IC 50 levels of ADR, Taxel and VCR is reduced in MCF-7/ADM and KPL-4/ADM cells after plasmid delivery

Article Snippet: The detection of WNT2 in cell culture medium was performed according to the manufacture’s protocols provided by Human Protein WNT2 ELISA kit (CSB-EL026133HU, Sino-American Biotechnology Co., Ltd., Hunan, Hubei, China).

Techniques: Plasmid Preparation, Control