Journal: Clinical Medicine. Oncology

Article Title: Disruption of the Non-Canonical WNT Pathway in Lung Squamous Cell Carcinoma

doi:

Figure Lengend Snippet: Schematic representation of the canonical and non-canonical Wnt pathways. sFRPs are inhibitors of both the canonical and non-canonical branches of the Wnt pathway. ( a ) Canonical Wnt pathway in its off state. ( b ) Canonical Wnt pathway in its on state. ( c ) Non-canonical Wnt pathway. Color halos represent genes that were used in this study. Grey: SFRP1, SFRP2, SFRP3, SFRP4, SFRP5; Blue: WNT1, WNT3A; Purple: FZD1; Yellow: LRP5, LRP6; Red: CTNNB1; Orange: WNT5A, WNT11; Teal: FZD2, FZD3, FZD6; Green: DVL2.

Article Snippet: For quantitative PCR, TaqMan primers (primer IDs in parentheses) for FZD3 (Hs00184043_m1), DVL2 (Hs00182901_m1), and CTNNB1 (Hs00170025_m1) were purchased from Applied Biosystems (Applied Biosystems, CA, U.S.A.).

Techniques:

Journal: Clinical Medicine. Oncology

Article Title: Disruption of the Non-Canonical WNT Pathway in Lung Squamous Cell Carcinoma

doi:

Figure Lengend Snippet: Differential expression of FZD3 , DVL2 and CTNNB1 between 10 lung squamous cell carcinoma and matched normals (Samples T21–T30). Results are generated by real-time RT-PCR using TaqMan gene specific primers from Applied Biosystems.

Article Snippet: For quantitative PCR, TaqMan primers (primer IDs in parentheses) for FZD3 (Hs00184043_m1), DVL2 (Hs00182901_m1), and CTNNB1 (Hs00170025_m1) were purchased from Applied Biosystems (Applied Biosystems, CA, U.S.A.).

Techniques: Quantitative Proteomics, Generated, Quantitative RT-PCR

Journal: Clinical Medicine. Oncology

Article Title: Disruption of the Non-Canonical WNT Pathway in Lung Squamous Cell Carcinoma

doi:

Figure Lengend Snippet: Pairwise expression correlation of genes in WNT pathway.

Article Snippet: For quantitative PCR, TaqMan primers (primer IDs in parentheses) for FZD3 (Hs00184043_m1), DVL2 (Hs00182901_m1), and CTNNB1 (Hs00170025_m1) were purchased from Applied Biosystems (Applied Biosystems, CA, U.S.A.).

Techniques: Expressing

Journal: Clinical Medicine. Oncology

Article Title: Disruption of the Non-Canonical WNT Pathway in Lung Squamous Cell Carcinoma

doi:

Figure Lengend Snippet: Primer sequences and conditions for RT-PCR analysis.

Article Snippet: For quantitative PCR, TaqMan primers (primer IDs in parentheses) for FZD3 (Hs00184043_m1), DVL2 (Hs00182901_m1), and CTNNB1 (Hs00170025_m1) were purchased from Applied Biosystems (Applied Biosystems, CA, U.S.A.).

Techniques: Sequencing

Journal: BMC biology

Article Title: DIX domain containing 1 (DIXDC1) modulates VEGFR2 level in vasculatures to regulate embryonic and postnatal retina angiogenesis.

doi: 10.1186/s12915-022-01240-3

Figure Lengend Snippet: Fig. 5 DIXDC1 upregulates VEGFR2 via Dvl2. A DIXDC1 or control vector, and VEGFR2 was co-transfected in HEK293T. Immunoprecipitation with antibody against DIXDC1 and VEGFR2 result revealed that no direct interaction between DIXDC1 and VEGFR2. B DIXDC1 or control vector, and Dvl2 was co-transfected in HEK293T. Immunoprecipitation with antibody against DIXDC1 and Dvl2 result revealed that there is an interaction between DIXDC1 and Dvl2. C Endogenous immunoprecipitation with antibodies against Dvl2 and VEGFR2 and there is an interaction between the two proteins in HUVEC. D Control siRNA or siDIX1 is transfected to HUVEC and subjected to immunoprecipitation antibody against Dvl2. E, F, G, and H Quantification of relative intensity of immunoprecipitated VEGFR2, Dvl2, and input VEGFR2 and Dvl2 of D. I DIXDC1 was overexpressed and there was a positive correlation between the expression level of DIXDC1 and Dvl2. J Quantification of relative intensity of Dvl2 in I. K DIXDC1, Dvl2, and VEGFR2 were co-overexpressed in HEK293T and the expression level of VEGFR2 and Dvl2 was observed. L Quantification of relative intensity of VEGFR2 in K. All Experiments were repeated at least 4 different sets. *p < 0.05, **p < 0.005, and ***p < 0.0001, by paired, 2-tailed Student’s t test and one-way ANOVA. Error bars represent the mean ± SD. Individual values can be found in Additional file 6: Fig 5

Article Snippet: The following primary antibodies were used: CD31(BD Pharmingen, Catalog 550274); DIXDC1 (Santa Cruz Biotechnology, Catalog sc-377160); VEGFR2 (Cell Signaling Technology, Catalog 9572); Dvl2 (Cell Signaling Technology, Catalog 3216); BrdU (Santa Cruz Biotechnology, Catalog sc-32323); β-catenin (Santa Cruz Biotechnology, Catalog sc-59737); Non-phospho- β-catenin (Cell Signaling Technology, Catalog 19807); DAPI (Sigma-Aldrich, Catalog D9542); β-actin (Thermo Fisher Scientific, Catalog MA5-15739).

Techniques: Control, Plasmid Preparation, Transfection, Immunoprecipitation, Expressing

Journal: BMC biology

Article Title: DIX domain containing 1 (DIXDC1) modulates VEGFR2 level in vasculatures to regulate embryonic and postnatal retina angiogenesis.

doi: 10.1186/s12915-022-01240-3

Figure Lengend Snippet: Fig. 6 DIXDC1 increases wnt/ β-catenin signaling and VEGFR2 stability via Dvl2. A knockdown of DIXDC1 affects the VEGF-induced signaling pathways (40 ng/ml) in HUVEC. B, C, and D Quantification of relative intensity of p-VEGFR2(Y1175), P-ERK1/2, and p-FAK of A. E Knockdown of DIXDC1 affects the wnt/β-catenin signaling pathways (200 ng/ml wnt3a) in HUVEC. F, G, H, and I Quantification of relative intensity of VEGFR2, non-p-β-catenin, β-catenin, and Dvl2 of E. J Interaction between Dvl2 and VEGFR2 is affected by DIXDC1 and wnt3a (200 ng/ml) in HUVEC. K, L Quantification of relative intensity of immunoprecipitated VEGFR2 and Dvl2. M Suppression of DIXDC1 affects the level of HIF1a in oxygen glucose-deprived condition in HUVEC. N, O, P, Q, R, and S Quantification of relative intensity of VEGFR2, HIF1a, non-p-β-catenin, β-catenin, Dvl2, and DIXDC1 of M. T VEGFR2, Dvl2, and DIXDC1 transfected to HEK293T and treated with MG132. Accumulation of VEGFR2 level was observed with MG132. U Quantification of relative intensity of VEGFR2 of T. All Experiments were repeated at least 4 different sets. *p < 0.05, **p < 0.005, and ***p < 0.0001, by one-way ANOVA. Error bars represent the mean ± SD. Individual values can be found in Additional file 6: Fig. 6

Article Snippet: The following primary antibodies were used: CD31(BD Pharmingen, Catalog 550274); DIXDC1 (Santa Cruz Biotechnology, Catalog sc-377160); VEGFR2 (Cell Signaling Technology, Catalog 9572); Dvl2 (Cell Signaling Technology, Catalog 3216); BrdU (Santa Cruz Biotechnology, Catalog sc-32323); β-catenin (Santa Cruz Biotechnology, Catalog sc-59737); Non-phospho- β-catenin (Cell Signaling Technology, Catalog 19807); DAPI (Sigma-Aldrich, Catalog D9542); β-actin (Thermo Fisher Scientific, Catalog MA5-15739).

Techniques: Knockdown, Protein-Protein interactions, Immunoprecipitation, Transfection

Journal: PLOS ONE

Article Title: Regulation of YAP and Wnt signaling by the endosomal protein MAMDC4

doi: 10.1371/journal.pone.0296003

Figure Lengend Snippet: (A) HEK293 cells over-expressing MAMDC4 were stimulated with control or Wnt3a conditioned media. Lysates were immunoprecipitated with antibodies against MAMDC4 or control IgG. MAMDC4 associates with LRP6, p-LRP6, total β-catenin, DVL2 and Axin in control and Wnt3a stimulated conditions. There is a substantial amount of LRP6 and p-LRP6 (relative to input) associated with MAMDC4 in both Wnt stimulated and unstimulated conditions. (B) Quantification of signal from immunoprecipitation relative to input. LRP6 and p-LRP6 have a ratio of approximately 1 while the ratio of β-catenin and Axin1 have a ratio of IP to input that is <1/10 th of the amount of LRP6 and p-LRP6 immunoprecipitated. All immunoprecipitations were repeated 3 to 5 times and the input lanes represents 1/50 th of lysate used for the immunoprecipitation.

Article Snippet: Cells were transfected with 3XFLAG (WT) DVL2 (cat. 24803; addgene, [ ]) and MAMDC4 (Wilson Lab) or Flag-Axin1 (cat. 109370; addgene, [ ] and MAMDC4 (Wilson Lab) using Lipofectamine LTX (cat. 15338030; ThermoFisher) following the manufacturer’s instructions.

Techniques: Expressing, Control, Immunoprecipitation

Journal: PLOS ONE

Article Title: Regulation of YAP and Wnt signaling by the endosomal protein MAMDC4

doi: 10.1371/journal.pone.0296003

Figure Lengend Snippet: (A) SKCO cells were transfected with MAMDC4 and DVL2. 48 hours post transfection cultures were treated with vehicle or Wnt3a for 1hr. Cells were labeled with antibodies to MAMDC4 (green) and DVL2 (red) and counterstained with DAPI. MAMDC4 and DVL2 are colocalized in puncta regardless of Wnt3a stimulation Scale bar : 10μm. (B) Quantification of Pearson’s Correlation Coefficient (r). Wnt3a treatment does not influence MAMDC4 and DVL2 colocalization. (C) SKCO cells were transfected with MAMDC4 and Axin1. 48 hours post transfection cultures were treated with vehicle or Wnt3a for 5min. Cells were labeled with antibodies to MAMDC4 (green) and Axin1 (red) and counterstained with DAPI. MAMDC4 and Axin1 are colocalized in some puncta after Wnt3a stimulation (arrows). (D) Quantification of Pearson’s Correlation Coefficient (r). There is an increase in MAMDC4 and Axin1 colocalization that is dependent on Wnt3a stimulation. (E) Immunoblot of NeutrAvidin Pull down. (F) Quantification of LRP6 Pull down / Lysate ratio.

Article Snippet: Cells were transfected with 3XFLAG (WT) DVL2 (cat. 24803; addgene, [ ]) and MAMDC4 (Wilson Lab) or Flag-Axin1 (cat. 109370; addgene, [ ] and MAMDC4 (Wilson Lab) using Lipofectamine LTX (cat. 15338030; ThermoFisher) following the manufacturer’s instructions.

Techniques: Transfection, Labeling, Western Blot

Journal: PLOS ONE

Article Title: Regulation of YAP and Wnt signaling by the endosomal protein MAMDC4

doi: 10.1371/journal.pone.0296003

Figure Lengend Snippet: SKCO cells were co-transfected with either MAMDC4 or MAMDC4 mutants and DVL2. 48 hours post transfection cells were labeled with antibodies to MAMDC4 (green) and DVL2 (red) and counterstained with DAPI. (A) MAMDC4 and DVL2 are colocalized in puncta, however, DVL2 has diffuse labeling in cells lacking MAMDC4 overexpression (arrow) (B) Quantification of number of cells expressing both MAMDC4 and DVL2, which contain DVL2 puncta. 35 to 45 cells were assessed for each condition, n = 3. **P<0.001, ***P<0.001. Statistical significance was determined using an Ordinary one-way ANOVA. Scale bar : 10μm.

Article Snippet: Cells were transfected with 3XFLAG (WT) DVL2 (cat. 24803; addgene, [ ]) and MAMDC4 (Wilson Lab) or Flag-Axin1 (cat. 109370; addgene, [ ] and MAMDC4 (Wilson Lab) using Lipofectamine LTX (cat. 15338030; ThermoFisher) following the manufacturer’s instructions.

Techniques: Transfection, Labeling, Over Expression, Expressing

Journal: Brain Sciences

Article Title: Expression of WNT Signaling Genes in the Dorsolateral Prefrontal Cortex in Schizophrenia

doi: 10.3390/brainsci14070649

Figure Lengend Snippet: TaqMan primers.

Article Snippet: DVL2 , Disheveled segment polarity protein 2 , Hs01005253_m1.

Techniques: Ubiquitin Proteomics

Journal: Brain Sciences

Article Title: Expression of WNT Signaling Genes in the Dorsolateral Prefrontal Cortex in Schizophrenia

doi: 10.3390/brainsci14070649

Figure Lengend Snippet: Relative mRNA expression of WNT signaling pathway genes in the dorsolateral prefrontal cortex (DLPFC) tissue of schizophrenia ( n = 20) compared to non-psychiatrically ill control ( n = 20) subjects, reported as percent of control (geometric mean of reference genes β2M and GAPDH). Data presented as mean ± standard error of the mean (SEM). Abbreviations : IQGAP1, IQ motif containing GTPase activating protein 1; CTNNβ1, Beta-catenin; GSK3β, Glycogen synthase kinase 3 beta; FOXO1, Forkhead box O1; LRP6, Lipoprotein receptor-related protein 6; MGEA5, Meningioma expressed antigen 5; TCF4, Transcription factor 4; βTRC, Beta-transducin repeat containing E3 ubiquitin protein ligase; PPP1Cβ, Protein phosphatase 1 catalytic subunit beta; DVL2, Disheveled segment polarity protein 2.

Article Snippet: DVL2 , Disheveled segment polarity protein 2 , Hs01005253_m1.

Techniques: Expressing, Control, Ubiquitin Proteomics

Journal: Brain Sciences

Article Title: Expression of WNT Signaling Genes in the Dorsolateral Prefrontal Cortex in Schizophrenia

doi: 10.3390/brainsci14070649

Figure Lengend Snippet: Heatmap generated from Kaleidoscope “lookup” study assessing mRNA expression of WNT signaling pathway genes across 20 RNA-seq and microarray-derived brain region-level transcriptomic datasets between schizophrenia and control groups. The heatmap utilizes unsupervised hierarchical clustering for both genes and datasets. The data are shown as log 2 -fold change (LFC) values: red indicates increased mRNA expression and blue indicates decreased mRNA expression on a scale of 2 to −2. The x-axis lists the WNT genes queried, and the y-axis lists the Kaleidoscope datasets. Abbreviations : IQGAP1, IQ motif containing GTPase activating protein 1; CTNNβ1, Beta-catenin; GSK3β, Glycogen synthase kinase 3 beta; FOXO1, Forkhead box O1; LRP6, Lipoprotein receptor-related protein 6; TCF4, Transcription factor 4; βTRC, Beta-transducin repeat containing E3 ubiquitin protein ligase; PPP1Cβ, Protein phosphatase 1 catalytic subunit beta; DVL2, Disheveled segment polarity protein 2.

Article Snippet: DVL2 , Disheveled segment polarity protein 2 , Hs01005253_m1.

Techniques: Generated, Expressing, RNA Sequencing, Microarray, Derivative Assay, Control, Ubiquitin Proteomics

Journal: Brain Sciences

Article Title: Expression of WNT Signaling Genes in the Dorsolateral Prefrontal Cortex in Schizophrenia

doi: 10.3390/brainsci14070649

Figure Lengend Snippet: Heatmap generated from Kaleidoscope “lookup” study assessing mRNA expression of WNT signaling pathway genes across 15 RNA-seq and microarray-derived cell-level transcriptomic datasets between schizophrenia and control groups. The heatmap utilizes unsupervised hierarchical clustering for both genes and datasets. The data are shown as log 2 -fold change (LFC) values: red indicates increased mRNA expression and blue indicates decreased mRNA expression on a scale of 2 to −2. The x-axis lists the WNT genes queried, and the y-axis lists the Kaleidoscope datasets. Abbreviations : IQGAP1, IQ motif containing GTPase activating protein 1; CTNNβ1, Beta-catenin; GSK3β, Glycogen synthase kinase 3 beta; FOXO1, Forkhead box O1; LRP6, Lipoprotein receptor-related protein 6; TCF4, Transcription factor 4; βTRC, Beta-transducin repeat containing E3 ubiquitin protein ligase; PPP1Cβ, Protein phosphatase 1 catalytic subunit beta; DVL2, Disheveled segment polarity protein 2.

Article Snippet: DVL2 , Disheveled segment polarity protein 2 , Hs01005253_m1.

Techniques: Generated, Expressing, RNA Sequencing, Microarray, Derivative Assay, Control, Ubiquitin Proteomics

Journal: Brain Sciences

Article Title: Expression of WNT Signaling Genes in the Dorsolateral Prefrontal Cortex in Schizophrenia

doi: 10.3390/brainsci14070649

Figure Lengend Snippet: Heatmap generated from Kaleidoscope “lookup” study assessing mRNA expression of WNT signaling pathway genes across 15 RNA-seq and microarray rodent-model derived transcriptomic datasets between antipsychotic-treated and vehicle-treated (control) groups. The heatmap utilizes unsupervised hierarchical clustering for both genes and datasets. The data are shown as log 2 -fold change (LFC) values: red indicates increased mRNA expression and blue indicates decreased mRNA expression on a scale of 2 to −2. The x-axis lists the WNT genes queried, and the y-axis lists the Kaleidoscope datasets. Abbreviations : IQGAP1, IQ motif containing GTPase activating protein 1; CTNNβ1, Beta-catenin; GSK3β, Glycogen synthase kinase 3 beta; FOXO1, Forkhead box O1; LRP6, Lipoprotein receptor-related protein 6; TCF4, Transcription factor 4; βTRC, Beta-transducin repeat containing E3 ubiquitin protein ligase; PPP1Cβ, Protein phosphatase 1 catalytic subunit beta; DVL2, Disheveled segment polarity protein 2.

Article Snippet: DVL2 , Disheveled segment polarity protein 2 , Hs01005253_m1.

Techniques: Generated, Expressing, RNA Sequencing, Microarray, Derivative Assay, Control, Ubiquitin Proteomics