rabbit polyclonal anti dvl1  (Proteintech)

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Levels of the 7TGP fluorescent Wnt reporter in HEK293T cells transfected with control siRNA, Dvl1-specific siRNA, Nup358-specific siRNA, or a combination of Nup358-specific siRNA and Dvl1-specific siRNA and treated with either Wnt3a or vehicle. b. Western blot analysis and relative quantification of protein levels of Dvl1 and Axin1 in HEK293T cells transfected with either control or Nup358-specific siRNA. α-Tubulin and GAPDH were used as loading controls. Data are expressed as mean ± SD. **p ≤ 0.01. c. Immunofluorescence staining for Dvl1 and Nup358 in HEK293T cells transfected with either control or Nup358-specific siRNA. d. Immunofluorescence staining for Dvl1 and Nup358 in HEK293T cells transfected with control or Nup358-specific siRNA and treated with vehicle or 1,6-hexanediol 5% for 2 minutes. e. Fluorescence recovery after photobleaching (FRAP) and fusion analysis of Dvl1 condensates in HEK293T cells transfected with Nup358-specific siRNA and EGFP-Dvl1.

Article Snippet: Precleared protein solution was incubated over-night at 4°C on a rotator with either a specific anti-Dvl1 antibody (Santa Cruz #8025) at the final dilution of 1:400 or with the same amount of control rabbit IgG.

Techniques: Transfection, Control, Western Blot, Quantitative Proteomics, Immunofluorescence, Staining, Fluorescence

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. HEK293T cells were transfected with β-catenin-GFP plasmid and control or Nup358-specific siRNAs and GFP expression was analyzed by confocal microscopy. The percentage of GFP-positive cells and GFP intensity per cell were quantified. Data are expressed as mean ± SD. ***p ≤ 0.001. b. Immunofluorescence staining for Dvl1 and ADP-ribosylation in HEK293T cells transfected with either control or Nup358-specific siRNAs. c. Western blot analysis of co-immunoprecipitation of ADP-ribosylation and Axin1 in HEK293T cells transfected with control or Nup358-specific siRNA. Axin1 levels in immunoprecipitates were quantified relative to controls and normalized to Axin1 input levels. Data are expressed as mean ± SD. **p ≤ 0.01 d. Left: Western blot analyses of Nup358 and Axin1 in HEK293T cells transfected with control or Nup358-specific siRNA and treated with 200 nM XAV-939 or vehicle for 72 hours. α-Tubulin was used as loading control. Right: Quantification of Axin1 protein levels (n=2 experiments). e. Quantification of fluorescent Wnt reporter activity by flow cytometry analysis in HEK293T cells transfected with control siRNA or Nup358-specific siRNA and treated with 200nM XAV-939 or vehicle for 72 hours. Data are expressed as mean ± SD. **p ≤ 0.01 and ***p ≤ 0.001.

Article Snippet: Precleared protein solution was incubated over-night at 4°C on a rotator with either a specific anti-Dvl1 antibody (Santa Cruz #8025) at the final dilution of 1:400 or with the same amount of control rabbit IgG.

Techniques: Transfection, Plasmid Preparation, Control, Expressing, Confocal Microscopy, Immunofluorescence, Staining, Western Blot, Immunoprecipitation, Activity Assay, Flow Cytometry

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Schematic representing different functional domains of Nup358. b. Fluorescent Wnt reporter activity in HEK293T cells transfected with control siRNA, Nup358-specific siRNA, Ubc9-specific siRNA, or treated with the SUMOylation inhibitor 2-D08. c. Schematic representing different HA-tagged truncated forms of Nup358 that were transiently expressed in HEK293T cells. d. Fluorescent Wnt reporter activity in HEK293T cells transfected with either control or Nup358-specific siRNA alone or in combination with different HA-tagged truncated forms of Nup358 were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. ****p ≤ 0.0001 e. Western blot analysis of co-immunoprecipitation of HA-Nup358 and EGFP-Dvl1 transiently expressed in HEK293T cells. GAPDH was used as loading control. f. Immunofluorescence of HEK293T cells transfected with HA-Tagged Nup358 1–1133 and EGFP-Dvl1and stained with an anti-HA antibody. g. Immunofluorescence staining for endogenous Dvl1 and Nup358 in HEK293T cells transfected with either scramble control or Nup358-specific siRNA alone or in combination with HA-Tagged Nup358 1–1133 fragment were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. **p ≤ 0.01.

Article Snippet: Precleared protein solution was incubated over-night at 4°C on a rotator with either a specific anti-Dvl1 antibody (Santa Cruz #8025) at the final dilution of 1:400 or with the same amount of control rabbit IgG.

Techniques: Functional Assay, Activity Assay, Transfection, Control, Confocal Microscopy, Western Blot, Immunoprecipitation, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Levels of the 7TGP fluorescent Wnt reporter in HEK293T cells transfected with control siRNA, Dvl1-specific siRNA, Nup358-specific siRNA, or a combination of Nup358-specific siRNA and Dvl1-specific siRNA and treated with either Wnt3a or vehicle. b. Western blot analysis and relative quantification of protein levels of Dvl1 and Axin1 in HEK293T cells transfected with either control or Nup358-specific siRNA. α-Tubulin and GAPDH were used as loading controls. Data are expressed as mean ± SD. **p ≤ 0.01. c. Immunofluorescence staining for Dvl1 and Nup358 in HEK293T cells transfected with either control or Nup358-specific siRNA. d. Immunofluorescence staining for Dvl1 and Nup358 in HEK293T cells transfected with control or Nup358-specific siRNA and treated with vehicle or 1,6-hexanediol 5% for 2 minutes. e. Fluorescence recovery after photobleaching (FRAP) and fusion analysis of Dvl1 condensates in HEK293T cells transfected with Nup358-specific siRNA and EGFP-Dvl1.

Article Snippet: Plasmids used in this study include β-catenin-EGFP (Addgene, #71367), EGFP-Dvl1 (Addgene, #194586) and plasmids HA-Nup358 1–2683 , HA-Nup358 1–2148 , HA-Nup358 1–1810 , HA-Nup358 1–1133 from Wälde et al 13 .

Techniques: Transfection, Control, Western Blot, Quantitative Proteomics, Immunofluorescence, Staining, Fluorescence

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Schematic representing different functional domains of Nup358. b. Fluorescent Wnt reporter activity in HEK293T cells transfected with control siRNA, Nup358-specific siRNA, Ubc9-specific siRNA, or treated with the SUMOylation inhibitor 2-D08. c. Schematic representing different HA-tagged truncated forms of Nup358 that were transiently expressed in HEK293T cells. d. Fluorescent Wnt reporter activity in HEK293T cells transfected with either control or Nup358-specific siRNA alone or in combination with different HA-tagged truncated forms of Nup358 were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. ****p ≤ 0.0001 e. Western blot analysis of co-immunoprecipitation of HA-Nup358 and EGFP-Dvl1 transiently expressed in HEK293T cells. GAPDH was used as loading control. f. Immunofluorescence of HEK293T cells transfected with HA-Tagged Nup358 1–1133 and EGFP-Dvl1and stained with an anti-HA antibody. g. Immunofluorescence staining for endogenous Dvl1 and Nup358 in HEK293T cells transfected with either scramble control or Nup358-specific siRNA alone or in combination with HA-Tagged Nup358 1–1133 fragment were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. **p ≤ 0.01.

Article Snippet: Plasmids used in this study include β-catenin-EGFP (Addgene, #71367), EGFP-Dvl1 (Addgene, #194586) and plasmids HA-Nup358 1–2683 , HA-Nup358 1–2148 , HA-Nup358 1–1810 , HA-Nup358 1–1133 from Wälde et al 13 .

Techniques: Functional Assay, Activity Assay, Transfection, Control, Confocal Microscopy, Western Blot, Immunoprecipitation, Immunofluorescence, Staining

Journal: Biomolecules

Article Title: Melatonin as an Alleviator in Decabromodiphenyl Ether-Induced Aberrant Hippocampal Neurogenesis and Synaptogenesis: The Role of Wnt7a

doi: 10.3390/biom15081087

Figure Lengend Snippet: Wnt7a/FZD5-mediated canonical β-catenin-dependent genes thus promoted hippocampal neurogenesis in P21 rats. ( A – C ) Protein expression of Wnt3a, Wnt7a, DVL1, and downstream phos-GSK3β (Ser9) and total GSK3β. ( D , E ) Phosphorylated-β-catenin (Ser33/37/Thr41), total β-catenin, and nuclear translocation. ( F ) β-catenin target genes Prox1 , Neurod1 , and Neurog2 transcriptional level. ( G , H ) Protein expression of Prox1, NeuroD1, and Neurog2. GAPDH and PCNA were used for internal reference of total protein and nuclear protein, respectively. ( I ) Co-immunoprecipitation analysis of FZD5 binding to Wnt7a in the hippocampus of P21 rats. ( J ) Immunofluorescence labeling for Wnt7a (Mu, Red), NeuroD1 (Rb, Green), and DAPI (blue) in hippocampal DG at P21 rats. Scale bar: upper—50 μm; lower—20 μm. Data are presented as the mean ± SD (n = 3–4 rats per group). * p < 0.05, ** p < 0.01 vs. the control; & p < 0.05, && p < 0.01 vs. 30 mg/kg group; # p < 0.05, ## p < 0.01 vs. 100 mg/kg BDE209 group. ns, no significance.

Article Snippet: Primary antibodies with dilution used were as follows: the phospho-β-catenin (Ser33/37/Thr41) antibody was from CST (#9561); rabbit polyclonal antibodies, β-catenin (1:5000, #51067-2-AP), PSD95 (1:2000), MAP2 (1:2000, #17490-1-AP), Prox1(1:1000, #11067-2-AP), and NeuroD1 (1:1000, #12081-1-AP) were from Proteintech (Wuhan, China); Neurog2 (#A19800), Netrin1 (#A16236), CRMP3 (#A17615), Dvl1 (#A10536), phospho-GSK3β-Ser9 (#AP0039), and GSK3β (#A2081, 1:1000) were from Abclonal (Wuhan, China); and Wnt3a (#sc-136163) was from Santa Cruz (Dallas, TX, USA).

Techniques: Expressing, Translocation Assay, Immunoprecipitation, Binding Assay, Immunofluorescence, Labeling, Control

Journal: Biomolecules

Article Title: Melatonin as an Alleviator in Decabromodiphenyl Ether-Induced Aberrant Hippocampal Neurogenesis and Synaptogenesis: The Role of Wnt7a

doi: 10.3390/biom15081087

Figure Lengend Snippet: Melatonin triggered Wnt7a/FZD5 signaling target genes thus promoted synaptogenesis of P35 BDE-209-exposed rats. ( A – C ) Protein expression of Wnt3a, Wnt7a, DVL1, and downstream phos-GSK3β (Ser9) and total GSK3β. ( D , E ) Phos-β-catenin (Ser33/37/Thr41), total β-catenin, and nuclear translocation. ( F ) Wnt7a target genes Dlg4 , Nertin1 , and Crmp3/Dpysl4 transcriptional level. ( G , H ) Protein expression of PSD95, Netrin1, CRMP3, and mature neurons’ marker MAP2. GAPDH and PCNA were used for internal reference of total protein and nuclear protein, respectively. ( I ) Co-immunoprecipitation analysis of FZD5 binding to Wnt7a in the hippocampus of P35 rats. ( J ) Immunofluorescence labeling for Wnt7a (Mu, Red), MAP2 (Rb, Green), and DAPI (blue) in hippocampal DG at P35 rats. Scale bar: upper—50 μm; lower—20 μm. Data are presented as the mean ± SD (n = 3–4 rats per group). * p < 0.05, ** p < 0.01 vs. the control; && p < 0.01 vs. 30 mg/kg group; # p < 0.05, ## p < 0.01 vs. 100 mg/kg BDE209 group. ns, no significance.

Article Snippet: Primary antibodies with dilution used were as follows: the phospho-β-catenin (Ser33/37/Thr41) antibody was from CST (#9561); rabbit polyclonal antibodies, β-catenin (1:5000, #51067-2-AP), PSD95 (1:2000), MAP2 (1:2000, #17490-1-AP), Prox1(1:1000, #11067-2-AP), and NeuroD1 (1:1000, #12081-1-AP) were from Proteintech (Wuhan, China); Neurog2 (#A19800), Netrin1 (#A16236), CRMP3 (#A17615), Dvl1 (#A10536), phospho-GSK3β-Ser9 (#AP0039), and GSK3β (#A2081, 1:1000) were from Abclonal (Wuhan, China); and Wnt3a (#sc-136163) was from Santa Cruz (Dallas, TX, USA).

Techniques: Expressing, Translocation Assay, Marker, Immunoprecipitation, Binding Assay, Immunofluorescence, Labeling, Control