goat anti mouse wnt5a polyclonal ab  (R&D Systems)

Journal: Molecular Psychiatry

Article Title: Inhibition of colony stimulating factor 1 receptor corrects maternal inflammation-induced microglial and synaptic dysfunction and behavioral abnormalities

doi: 10.1038/s41380-020-0671-2

Figure Lengend Snippet: a Gene ontology (GO) biological processes pathway analysis shows that MIA microglia increase synaptogenic functions while repopulated microglia recover homeostatic functions. Left (red): Top significantly enriched GO biological process terms increased by MIA and decreased by repopulation. Right (purple): Top significantly enriched GO biological process terms decreased by MIA and increased by repopulation. These GO findings were verified using GORILLA. b IPA of genes with differential expression in microglia between MIA versus Saline (RNA-seq data). Pathway analysis reveals MIA-induced upregulation of neuritogenic gene expression, specifically in developmental stages, based on activation z -score. Red denotes pathway activated in E17 MIA microglia. c Genes in “neuritogenesis/formation of cellular protrusions” function. Hierarchal clustering of gene sets based on relative expression values; red: high relative expression, blue: low relative expression. Cluster 1 represents genes increased in adult MIA microglia but reduced in MIA + MG-REP including Ctnnd2, Ncam2, and Ntrk2 . Cluster 2 represents genes increased in immature MIA microglia including Ncam2, Ntn, Ptn and Wnt5a . Cluster3 represents genes decreased in immature MIA microglia including Plau. In situ hybridization (ISH) and immunofluorescence of E17 Saline or MIA offspring in the cortical plate region. d mRNA of cellular protrusion/ neuritogenic genes ( Ctnnd2, Ncam2, Ntn, Ptn, and Wnt5a ) were detected by florescent-labeled antisense cRNA probes (red) but not by scramble cRNA probe (not detected: N.D.), and the sections were immunostained for IBA1 (green) and DAPI (blue). e The number of IBA1 + cells expressing the cellular protrusion/neuritogenic genes were quantified in the cortical plate region. n = (4–5/2) male mice/ litters per molecule for Saline and MIA, n = 3 for scramble control probe. * p < 0.05, ** p < 0.01, ns denotes no significance, by unpaired Student t test. Graphs indicate mean ± s.e.m. ELISA verification of selected RNA-seq molecules: CTNND2 ( f ), NCAM2 ( g ), NTRK2 ( h ), NTN ( i) , PTN ( j ) and WNT5A ( k ) in acutely isolated microglia. MIA increases protein expression of cellular protrusion/neuriotgenic molecules in microglia that were normalized via repopulation. n = (6/4, 6/3, 5/3, 6/3) female mice/litters for P60 Saline + CTRL, MIA + CTRL, Saline + MG-REP and MIA + MG-REP. CTNND2: Prenatal treatment effect, F (1,19) = 157.1, p < 0.0001, Drug effect, F (1,19) = 262.7, p < 0.0001, Interaction effect, F (1,19) = 201, p < 0.0001, NCAM2: Prenatal treatment effect, F (1,19) = 23.76, p = 0.0001, Drug effect, F (1,19) = 26.29, p < 0.0001, Interaction effect, F (1,19) = 17.63, p = 0.0005, NTRK2: Prenatal treatment effect, F (1,18) = 13.99, p = 0.0015, Drug effect, F (1,18) = 12.45, p = 0.0024, Interaction effect, F (1,18) = 0.06203, p = 0.8061, NTN: Prenatal treatment effect, F (1,19) = 0.01669, p = 0.8986, Drug effect, F (1,19) = 0.8, p = 0.3823, Interaction effect, F (1,19) = 6.121, p = 0.0230, PTN: Prenatal treatment effect, F (1,19) = 10.31, p = 0.0046, Drug effect, F (1,19) = 52.02, p < 0.0001, Interaction effect, F (1,19) = 0.002927 p = 0.9574, WNT5A: Prenatal treatment effect, F (1,19) = 5.581, p = 0.0290, Drug effect, F (1,19) = 1.550, p = 0.2282, Interaction effect, F (1,19) = 0.0834 p = 0.7799, * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 as determined by 2-way ANOVA (alpha = 0.05) with Tukey’s post-hoc. # p < 0.05 for main effect of MIA. Graphs indicate mean ± s.e.m.

Article Snippet: For the detection of NCAM2, CTNND2 and WNT5A, custom ELISA kits were developed according to the manufacturer’s instruction using anti-NCAM2 goat polyclonal antibody (0.3 μg/well Acris Antibodies GmbH, AP32136PU-N), biotinylated anti-NCAM2 goat polyclonal antibody using Antibody Biotinylation Kit (0.3 μg/ml, Pierce/Thermo Scientific, 90407), anti-CTNND2 mouse monoclonal antibody (0.3 μg/well, Santa Cruz Biotechnology, SC-81793, clone 40.1), biotinylated anti-CTNND2 rabbit antibody (1 μg/ml, Abcam, EPR17628), anti-WNT5A goat polyclonal antibody (0.3 μg/well, R&D Systems, AF645), and biotinylated anti-WNT5A antibody using Antibody Biotinylation Kit (1 μg/ml, Pierce).

Techniques: Quantitative Proteomics, Saline, RNA Sequencing, Gene Expression, Activation Assay, Expressing, In Situ Hybridization, Immunofluorescence, Labeling, Control, Enzyme-linked Immunosorbent Assay, Isolation

Journal: International Journal of Oral-Medical Sciences

Article Title: <b>Wnt5a Stimulates the Bone Formation in Tension Side during Orthodontic Tooth Movement </b>

doi: 10.5466/ijoms.13.120

Figure Lengend Snippet: Fig.2 HE and immunohistochemical staining for wnt5a. After an orthodontic force was applied for 7 days, the arrangement of the fibers and fibroblasts was observed. It became coarse and irregular in the TF (10g) group (arrow heads), but not in the control group (A, B). Immunoreactivity for wnt5a was observed on the alveolar bone surface and in the PDL tissues (arrow heads) in the TF (10g) group (C, D). D: dentine, C: cementum, PDL: periodontal ligament, AB: alveolar bone. Original magnification 200×, Scale bar: 50μm

Article Snippet: After washing in Tris-buffered saline, the sections were incubated with polyclonal anti-goat wnt5a (R&D Systems Co., MN, USA) for 18 hrs at 4°C.

Techniques: Immunohistochemical staining, Staining, Control

Journal: International Journal of Oral-Medical Sciences

Article Title: <b>Wnt5a Stimulates the Bone Formation in Tension Side during Orthodontic Tooth Movement </b>

doi: 10.5466/ijoms.13.120

Figure Lengend Snippet: Fig.3 The number of Wnt5a-positive cells. The number of Wnt5a-positive cells was significantly different from the control (*p＜0.01).

Article Snippet: After washing in Tris-buffered saline, the sections were incubated with polyclonal anti-goat wnt5a (R&D Systems Co., MN, USA) for 18 hrs at 4°C.

Techniques: Control

Journal: International Journal of Oral-Medical Sciences

Article Title: <b>Wnt5a Stimulates the Bone Formation in Tension Side during Orthodontic Tooth Movement </b>

doi: 10.5466/ijoms.13.120

Figure Lengend Snippet: Fig.4 The time-course effect of wnt5a (100ng/ml) on the mRNA expression of ALP in hPDL cells. The hPDL cells were treated with wnt5a, then the mRNA expression of ALP was observed to increase in a time-dependent manner for up to 24h (Mann-Whitney U-test, *P<0.05, **P<0.01). The data shown are representative of four independent experiments using samples from six different patients.

Article Snippet: After washing in Tris-buffered saline, the sections were incubated with polyclonal anti-goat wnt5a (R&D Systems Co., MN, USA) for 18 hrs at 4°C.

Techniques: Expressing, MANN-WHITNEY

Journal: International Journal of Oral-Medical Sciences

Article Title: <b>Wnt5a Stimulates the Bone Formation in Tension Side during Orthodontic Tooth Movement </b>

doi: 10.5466/ijoms.13.120

Figure Lengend Snippet: Fig.5 The time-course effect of wnt5a (100ng/ml) on the mRNA expression of COL1 in hPDL cells. The hPDL cells were treated with wnt5a, then the mRNA expression of COL1 was observed to increase in a time-dependent manner for up to 24h (Mann-Whitney U-test, *P<0.05, **P<0.01). The data shown are representative of four independent experiments using samples from six different patients.

Article Snippet: After washing in Tris-buffered saline, the sections were incubated with polyclonal anti-goat wnt5a (R&D Systems Co., MN, USA) for 18 hrs at 4°C.

Techniques: Expressing, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Thyroid Hormone-Regulated Wnt5a/Ror2 Signaling Is Essential for Dedifferentiation of Larval Epithelial Cells into Adult Stem Cells in the Xenopus laevis Intestine

doi: 10.1371/journal.pone.0107611

Figure Lengend Snippet: Cross sections were immunostained with anti-Wnt5a (A–C), anti-Fzd2 (D–F), or anti-Ror2 (G–I). A, D, G: Stage 57. Cells positive for Ror2 (G; arrows) are scattered only in the larval epithelium (LE). They possess the brush border (bb) on the apical surface (Inset). B, E, H: Stage 61. Cells positive for Wnt5a (B) and Fzd2 (E) are broadly distributed in every tissue except for the degenerating larval epithelium, whereas a strong immunoreactivity for Ror2 is localized in islets of the adult epithelium (AE) (H). C, F, I: Stage 66. Immunoreactivity for each protein becomes weaker. CT: connective tissue; M: muscles; Scale bars: 20 µm.

Article Snippet: The in vitro -translated (IVT) sample (5 μl) was analyzed by Western blotting using a goat polyclonal antibody against mouse Wnt5a (diluted 1∶500; R&D Systems, Minneapolis, MN, USA).

Techniques: Muscles

Journal: PLoS ONE

Article Title: Thyroid Hormone-Regulated Wnt5a/Ror2 Signaling Is Essential for Dedifferentiation of Larval Epithelial Cells into Adult Stem Cells in the Xenopus laevis Intestine

doi: 10.1371/journal.pone.0107611

Figure Lengend Snippet: Cross sections were stained with MG-PY (A, C) or immunostained with anti-Ror2 (B, D). A, B: Control intestines. Cells positive for Ror2 are scattered in the simple columnar epithelium (B; arrow). C, D: Intestines cultured with the addition of Wnt5a. Epithelial cells positive for Ror2 change in morphology (arrows) and often invaginate into the connective tissue (CT). Scale bars: 20 µm.

Article Snippet: The in vitro -translated (IVT) sample (5 μl) was analyzed by Western blotting using a goat polyclonal antibody against mouse Wnt5a (diluted 1∶500; R&D Systems, Minneapolis, MN, USA).

Techniques: Staining, Control, Cell Culture

Journal: PLoS ONE

Article Title: Thyroid Hormone-Regulated Wnt5a/Ror2 Signaling Is Essential for Dedifferentiation of Larval Epithelial Cells into Adult Stem Cells in the Xenopus laevis Intestine

doi: 10.1371/journal.pone.0107611

Figure Lengend Snippet: Cross sections were double-immunostained with anti-Ror2 (green) and anti-Msi1 (A, C; red) or anti-PCNA (B, D; red) antibodies, and counterstained with DAPI. A, B: Control intestines. The simple columnar epithelium (E) remains negative for Msi1 (A). Proliferating cells positive for PCNA are very few, if any (B). C, D: Intestines cultured with the addition of Wnt5a. Epithelial cells positive for Ror2 invaginate into the connective tissue (CT) but remain negative for Msi1 (C). Proliferating cells are more numerous than those in the control intestines and are also detectable in cells negative for Ror2 (D; arrows). Scale bars: 20 µm.

Article Snippet: The in vitro -translated (IVT) sample (5 μl) was analyzed by Western blotting using a goat polyclonal antibody against mouse Wnt5a (diluted 1∶500; R&D Systems, Minneapolis, MN, USA).

Techniques: Control, Cell Culture

Journal: PLoS ONE

Article Title: Thyroid Hormone-Regulated Wnt5a/Ror2 Signaling Is Essential for Dedifferentiation of Larval Epithelial Cells into Adult Stem Cells in the Xenopus laevis Intestine

doi: 10.1371/journal.pone.0107611

Figure Lengend Snippet: Cross sections were stained with MG-PY (A, C) or immunostained with anti-Ror2 (B, D). A, B: Intestines treated with 20 nM T3. Adult stem/progenitor cells stained strongly red with MG-PY and positive for Ror2 (A, B; arrow) invaginate into the connective tissue (CT). C, D: T3-treated intestines cultured with the addition of anti-Wnt5a antibody. The epithelium (E) remains a single layer, where cells positive for Ror2 are scattered (D; arrows). Scale bars: 20 µm.

Article Snippet: The in vitro -translated (IVT) sample (5 μl) was analyzed by Western blotting using a goat polyclonal antibody against mouse Wnt5a (diluted 1∶500; R&D Systems, Minneapolis, MN, USA).

Techniques: Staining, Cell Culture

Journal: PLoS ONE

Article Title: Thyroid Hormone-Regulated Wnt5a/Ror2 Signaling Is Essential for Dedifferentiation of Larval Epithelial Cells into Adult Stem Cells in the Xenopus laevis Intestine

doi: 10.1371/journal.pone.0107611

Figure Lengend Snippet: Cross sections were double-immunostained with anti-Ror2 (green) and anti-Msi1 (A, C; red) or anti-PCNA (B, D; red) antibodies, and counterstained with DAPI. A, B: Intestines treated with 20 nM T3. Epithelial cells (E) invaginating into the connective tissue (CT) are double positive for Ror2 and Msi1 (A; arrow) and actively proliferate (B). C, D: T3-treated intestines cultured with the addition of anti-Wnt5a antibody. Epithelial cells positive for Ror2 remain negative for Msi1 (C). Proliferating cells are fewer than those in the T3-treated intestines in the absence of the antibody (D). Scale bars: 20 µm.

Article Snippet: The in vitro -translated (IVT) sample (5 μl) was analyzed by Western blotting using a goat polyclonal antibody against mouse Wnt5a (diluted 1∶500; R&D Systems, Minneapolis, MN, USA).

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: Thyroid Hormone-Regulated Wnt5a/Ror2 Signaling Is Essential for Dedifferentiation of Larval Epithelial Cells into Adult Stem Cells in the Xenopus laevis Intestine

doi: 10.1371/journal.pone.0107611

Figure Lengend Snippet: Larval absorptive cells expressing Ror2 are scattered in the simple columnar epithelium until stage 59. At stage 60, thyroid hormone up-regulates the expression of Wnt5a, whose protein binds to Ror2 of the absorptive cells and changes their morphology from simple columnar to roundish cells close to the connective tissue. This Wnt5a/Ror2 signaling is not sufficient but essential for epithelial dedifferentiation into the stem cells. In addition, Wnt5a promotes cell proliferation via receptors other than Ror2.

Article Snippet: The in vitro -translated (IVT) sample (5 μl) was analyzed by Western blotting using a goat polyclonal antibody against mouse Wnt5a (diluted 1∶500; R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing