rabbit anti mouse polyclonal ror2  (Vector Laboratories)

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: Cells expressing Ror2 in cardiac tissue was visualized in Ror2-LacZ mice 28 days after A) sham surgery or B) TAC surgery. C) Protein localization was visualized in cardiac tissue 7 days after TAC by IHC for Ror2. D) Ror1 and Ror2 protein expression was measured by western blot at various days after TAC surgery (β-actin as a loading control). E) Endothelial cells, leukocytes, and fibroblasts were isolated 7 days after Sham or TAC surgery, and RNA expression of relevant genes was quantified.

Article Snippet: Cardiac tissue sections were either stained for 5-bromo-4-chloro-3-indolyl-b-galactosidase (Xgal; Roche Cat #XGAL-RO) or immunostained with rabbit anti-mouse polyclonal Ror2 (provided by Dr. Yasuhiro Minami, Kobe University) with DAB visualization (Vector Laboratories, Cat #SK-4100), and counterstained with hematoxylin and eosin (Sigma Aldrich, Cat #HHS128-4L and #HT110180-2.5L, respectively).

Techniques: Expressing, Western Blot, Isolation, RNA Expression

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: Cells expressing Ror2 in cardiac tissue was visualized in Ror2-LacZ mice 28 days after A) sham surgery or B) TAC surgery. C) Protein localization was visualized in cardiac tissue 7 days after TAC by IHC for Ror2. D) Ror1 and Ror2 protein expression was measured by western blot at various days after TAC surgery (β-actin as a loading control). E) Endothelial cells, leukocytes, and fibroblasts were isolated 7 days after Sham or TAC surgery, and RNA expression of relevant genes was quantified.

Article Snippet: Protein lysates from TAC heart samples were isolated from cardiac tissue by 1% Triton-X supplemented with protease inhibitors and probed for rabbit anti-mouse polyclonal Ror1 (provided by Dr. Michael Greenberg, ) rabbit anti-mouse monoclonal Ror2 (Cell Signaling Technology Cat #88639), or β-actin (Cell Signaling Technology Cat #4970) by western blot.

Techniques: Expressing, Western Blot, Control, Isolation, RNA Expression

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: Murine cardiac fibroblasts were isolated from healthy cardiac tissue of C57Bl6 mice. A) RNA expression of genes associated with fibroblast identity, fibroblast activation, Ror1/2 receptors, and planar cell polarity signaling was quantified over time. Cardiac myofibroblasts at passage 2 were stained for SMØ-actin fibers after B) control and C) TGF-β1 treatment. D) RNA expression of genes associated with myofibroblast differentiation, inflammation, and Ror1/2 receptors was quantified in control and TGF-β1 treated fibroblasts.

Article Snippet: Protein lysates from TAC heart samples were isolated from cardiac tissue by 1% Triton-X supplemented with protease inhibitors and probed for rabbit anti-mouse polyclonal Ror1 (provided by Dr. Michael Greenberg, ) rabbit anti-mouse monoclonal Ror2 (Cell Signaling Technology Cat #88639), or β-actin (Cell Signaling Technology Cat #4970) by western blot.

Techniques: Isolation, RNA Expression, Activation Assay, Staining, Control

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: Primary cardiac fibroblasts were isolated from healthy control and transgenic Ror1/2 double-knockout mice transcriptional phenotype was assessed by bulk RNA sequencing. A) RNA transcript reads of specific genes related to planar cell polarity, inflammatory cytokines, proliferation and cell division, ERK1/2 signaling, matrix remodeling, natriuretic signaling, and myofibroblast differentiation were quantified as normalized Z-score (4 samples for each genotype). B) Gene ontology analysis of differential gene expression between control and Ror1/2-KO cardiac fibroblasts showed the top 10 up-regulated and down-regulated terms by q-value, with terms grouped by color: cell activation in orange, inflammation in red, proliferation in green, and microtubule regulation in black. C) Significance versus fold change of each gene between control and Ror1/2-KO samples was visualized by volcano plot, with genes in each gene ontology term highlighted in corresponding colors (all other genes in grey).

Article Snippet: Protein lysates from TAC heart samples were isolated from cardiac tissue by 1% Triton-X supplemented with protease inhibitors and probed for rabbit anti-mouse polyclonal Ror1 (provided by Dr. Michael Greenberg, ) rabbit anti-mouse monoclonal Ror2 (Cell Signaling Technology Cat #88639), or β-actin (Cell Signaling Technology Cat #4970) by western blot.

Techniques: Isolation, Control, Transgenic Assay, Double Knockout, RNA Sequencing, Gene Expression, Activation Assay

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: Myofibroblast differentiation was induced by treatment with 10ng/mL TGF-β1 for 4 days. A) RNA expression of myofibroblast-related (Acta2 and Postn) and inflammation-related (IL-6) genes was quantified in control and Ror1/2-KO fibroblasts. B) SMΓ]-actin filaments were visualized by immunofluorescent staining of sub-confluent cells in control and Ror1/2-KO fibroblasts, and C) alignment of SMΓ1-actin filaments was quantified and normalized per cell.

Article Snippet: Protein lysates from TAC heart samples were isolated from cardiac tissue by 1% Triton-X supplemented with protease inhibitors and probed for rabbit anti-mouse polyclonal Ror1 (provided by Dr. Michael Greenberg, ) rabbit anti-mouse monoclonal Ror2 (Cell Signaling Technology Cat #88639), or β-actin (Cell Signaling Technology Cat #4970) by western blot.

Techniques: RNA Expression, Control, Staining

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: A) Transgenic Ror1/2 double knockout and control mice were subjected to TAC surgery, and cardiac output was imaged by echocardiography. B) Cardiac output factors were quantified: fractional shortening, left ventricular posterior wall thickness at end diastole, left ventricular diameter at end systole, and left ventricular diameter at end diastole. C) Survival of Ror1/2 double knockout mice and control mice after TAC surgery was recorded each day.

Article Snippet: Protein lysates from TAC heart samples were isolated from cardiac tissue by 1% Triton-X supplemented with protease inhibitors and probed for rabbit anti-mouse polyclonal Ror1 (provided by Dr. Michael Greenberg, ) rabbit anti-mouse monoclonal Ror2 (Cell Signaling Technology Cat #88639), or β-actin (Cell Signaling Technology Cat #4970) by western blot.

Techniques: Transgenic Assay, Double Knockout, Control

Journal: bioRxiv

Article Title: The Cell Surface Receptors Ror1/2 Control Cardiac Myofibroblast Differentiation

doi: 10.1101/2021.03.02.433549

Figure Lengend Snippet: Inflammatory profile of control and transgenic Ror1/2 double knockout mice was assessed after TAC or sham surgery. A) H&E staining of cardiac tissues 3-days post-TAC were imaged. B) Cells were isolated from cardiac tissue and relative quantity of leukocyte populations were determined by flow cytometry, C) quantified by absolute number. D) Gene expression of pro-inflammatory cytokines in cardiac lysate was measured. E) Vascular permeability at 1-day post-TAC was assessed by Evans Blue dye injection, with vascular leakage visualized by blue dye in the cardiac tissue.

Article Snippet: Protein lysates from TAC heart samples were isolated from cardiac tissue by 1% Triton-X supplemented with protease inhibitors and probed for rabbit anti-mouse polyclonal Ror1 (provided by Dr. Michael Greenberg, ) rabbit anti-mouse monoclonal Ror2 (Cell Signaling Technology Cat #88639), or β-actin (Cell Signaling Technology Cat #4970) by western blot.

Techniques: Control, Transgenic Assay, Double Knockout, Staining, Isolation, Flow Cytometry, Gene Expression, Permeability, Injection