Journal: Translational Oncology

Article Title: Distinct Patterns of Stromal and Tumor Expression of ROR1 and ROR2 in Histological Subtypes of Epithelial Ovarian Cancer 1

doi: 10.1016/j.tranon.2017.01.014

Figure Lengend Snippet: ROR expression in OC stroma. (A) Representative images of stromal ROR1 (top panel) and ROR2 (bottom panel) IHC, from a score of 0 to 3. ROR1 staining did not have strong expression in the stroma, and therefore, no representative figure of score 3 is shown (NA). (B) ROR expression in the stroma depicted by histological subtype. Each pie chart represents percentage of samples and ROR score per subtype. Scores of 0 to 3 colored white, light gray, gray, and dark gray, respectively.

Article Snippet: Sections were stained with Anti-ROR1 Ab (Abcam #135669) or Anti-ROR2 Ab (QED Biosciences, #34045) both at a 1:100 dilution in addition to a negative IgG control, and counterstained with hematoxylin.

Techniques: Expressing, Staining

Journal: Translational Oncology

Article Title: Distinct Patterns of Stromal and Tumor Expression of ROR1 and ROR2 in Histological Subtypes of Epithelial Ovarian Cancer 1

doi: 10.1016/j.tranon.2017.01.014

Figure Lengend Snippet: ROR expression in OC tumor. (A) Representative images of tumor ROR1 (top panel) and ROR2 (bottom panel) IHC, from a score of 0 to 3. (B) ROR expression in the tumor depicted by subtype. Each pie chart represents percentage of samples and ROR score per subtype. Scores of 0 to 3 colored white, light gray, gray, and dark gray, respectively.

Article Snippet: Sections were stained with Anti-ROR1 Ab (Abcam #135669) or Anti-ROR2 Ab (QED Biosciences, #34045) both at a 1:100 dilution in addition to a negative IgG control, and counterstained with hematoxylin.

Techniques: Expressing

Journal: Translational Oncology

Article Title: Distinct Patterns of Stromal and Tumor Expression of ROR1 and ROR2 in Histological Subtypes of Epithelial Ovarian Cancer 1

doi: 10.1016/j.tranon.2017.01.014

Figure Lengend Snippet: ROR expression relationship in tumor and stroma. Each panel represents number of tumor ROR expression scores for each stroma score ( x -axis, ROR S_0, ROR S_1, ROR S_2 and ROR S_3). Tumor ROR scores are represented by shading and distribution listed underneath. Each graph is paired with a representative image of the overall pattern of expression for that subtype. (A) ROR1 tumor and stroma expression in clear cell OC showed overall low stroma intensity but high tumor intensity. (B) ROR2 expression was overall low in both tumor and stroma in clear cell OC. (C) ROR1 expression in endometrioid OC was overall strong in tumor cells and weak in stroma. (D) ROR2 expression in endometrioid OC was very strong in tumor cells and absent in stroma. (E) ROR1 expression in mucinous OC was high in tumor and low in stroma. (F) ROR2 expression in mucinous OC was overall weak in both tumor and stroma. (G) ROR1 expression in serous tumor increased as stroma expression increased, but overall expression was mid to low. (H) ROR2 expression in serous OC tumor decreased as stroma expression increased. All cases with a stromal score of 3 exhibited absent tumor expression.

Article Snippet: Sections were stained with Anti-ROR1 Ab (Abcam #135669) or Anti-ROR2 Ab (QED Biosciences, #34045) both at a 1:100 dilution in addition to a negative IgG control, and counterstained with hematoxylin.

Techniques: Expressing

Journal: Translational Oncology

Article Title: Distinct Patterns of Stromal and Tumor Expression of ROR1 and ROR2 in Histological Subtypes of Epithelial Ovarian Cancer 1

doi: 10.1016/j.tranon.2017.01.014

Figure Lengend Snippet: Survival proportions of ROR stroma expression. (A): Kaplan-Meier curve representing PFS of patients (serous subtype only) with both ROR1 and ROR2 stroma positivity (red) and ROR1 and ROR2 stromal absence (blue). (B) Kaplan Meier curve representing OS of patients (serous subtype only) with both ROR1 and ROR2 stroma positivity (red) and ROR1 and ROR2 stromal absence (blue).

Article Snippet: Sections were stained with Anti-ROR1 Ab (Abcam #135669) or Anti-ROR2 Ab (QED Biosciences, #34045) both at a 1:100 dilution in addition to a negative IgG control, and counterstained with hematoxylin.

Techniques: Expressing

Journal: Translational Oncology

Article Title: Distinct Patterns of Stromal and Tumor Expression of ROR1 and ROR2 in Histological Subtypes of Epithelial Ovarian Cancer 1

doi: 10.1016/j.tranon.2017.01.014

Figure Lengend Snippet: Tracking ROR expression in individual case studies. (A) ROR1 expression in epithelial/tumor cells in individual cases as indicated by colored dots (blue = normal, red = primary, green = metastatic samples). Y -axis represents ROR expression score, and x -axis represents individual patient. (B) ROR1 expression in stromal cells in individual cases as indicated by colored dots (blue = normal, red = primary, green = metastatic samples). Y -axis represents ROR expression score, and x -axis represents individual patient. (C): ROR2 expression in epithelial/tumor cells in individual cases as indicated by colored dots (blue = normal, red = primary, green = metastatic samples). Y -axis represents ROR expression score, and x -axis represents individual patient. (D) ROR2 expression in stromal cells in individual cases as indicated by indicated by colored dots (blue = normal, red = primary, green = metastatic samples). Y -axis represents ROR expression score, and x -axis represents individual patient. (E) Representative images of ROR1 (left panel) and ROR2 (right panel) in case studies, with noticeable ROR2 expression in stromal components in primary and metastatic samples. (F) Representative image of case ID0681 recurrence showing diffuse and strong ROR2 expression in tumor and stroma.

Article Snippet: Sections were stained with Anti-ROR1 Ab (Abcam #135669) or Anti-ROR2 Ab (QED Biosciences, #34045) both at a 1:100 dilution in addition to a negative IgG control, and counterstained with hematoxylin.

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: Interdigital Hyperplasia in Holstein Cattle Is Associated With a Missense Mutation in the Signal Peptide Region of the Tyrosine-Protein Kinase Transmembrane Receptor Gene

doi: 10.3389/fgene.2019.01157

Figure Lengend Snippet: Manhattan plot of genome-wide association study for bovine interdigital hyperplasia. (A) Marker associations are plotted as negative log-transformed P values against the position in the bovine genome (UMD3.1.1). Two markers ARS-BFGL-NGS-64395 and ARS-BFGL-NGS-69582 exceed the genome-wide significance threshold of -log 10 P = 5.47 (black line). (B) Enlargement of the associated chromosomal region on BTA8 flanking markers ARS-BFGL-NGS-64395 and ARS-BFGL-NGS-69582. Genes located within this region, i.e., SPTLC1, ROR2, NFIL3, and AUH , are shown with black bars at their approximate positions.

Article Snippet: After blocking overnight at 4°C, immunoblots were incubated with primary anti-ROR2 antibody (1:500, ABIN2706970, Cohesion Biosciences, Aachen, Germany) and anti-β-actin (1:10,000, A5441, Sigma Aldrich, Darmstadt, Germany) at room temperature for 1 h. Incubation with the secondary antibodies (1:5000 for #1706515, 1:10,000 for #1706516, Bio-Rad, Munich, Germany) was done at room temperature for 1 h. Immunoblots were developed with Western ECL (GERPN2109, Sigma Aldrich, Darmstadt, Germany).

Techniques: GWAS, Marker, Transformation Assay, Genome Wide

Journal: Frontiers in Genetics

Article Title: Interdigital Hyperplasia in Holstein Cattle Is Associated With a Missense Mutation in the Signal Peptide Region of the Tyrosine-Protein Kinase Transmembrane Receptor Gene

doi: 10.3389/fgene.2019.01157

Figure Lengend Snippet: Genotype frequencies of the receptor tyrosine kinase-like orphan receptor 2 ( ROR2 ) variants rs377953295 (exon 1) and rs43572154 (exon 9) in interdigital hyperplasia (IH) type A ^a) affected and free (= healthy) cattle.

Article Snippet: After blocking overnight at 4°C, immunoblots were incubated with primary anti-ROR2 antibody (1:500, ABIN2706970, Cohesion Biosciences, Aachen, Germany) and anti-β-actin (1:10,000, A5441, Sigma Aldrich, Darmstadt, Germany) at room temperature for 1 h. Incubation with the secondary antibodies (1:5000 for #1706515, 1:10,000 for #1706516, Bio-Rad, Munich, Germany) was done at room temperature for 1 h. Immunoblots were developed with Western ECL (GERPN2109, Sigma Aldrich, Darmstadt, Germany).

Techniques:

Journal: Frontiers in Genetics

Article Title: Interdigital Hyperplasia in Holstein Cattle Is Associated With a Missense Mutation in the Signal Peptide Region of the Tyrosine-Protein Kinase Transmembrane Receptor Gene

doi: 10.3389/fgene.2019.01157

Figure Lengend Snippet: Detection of the receptor tyrosine kinase-like orphan receptor 2 ( ROR2 ) transcripts and isoforms in different bovine organs and tissues. (A) Organ and/or tissue samples (De, interdigital dermis, He, heart, Br, brain, Li, liver, Int, intestine, Ur, urinary bladder, Bo, bone marrow, Lu, lung, Sp, spleen, St, stomach) were collected from healthy cattle and RNA extracted. Relative expression levels were calculated using the average Δ Ct-values of three biological replicates with GAPDH as normalizer. (B) ROR2 isoform specific primers were used to amplify RNA from IntD, interdigital dermis, Lu, lung and Sp, spleen (NTC, non template control). Two amplicons corresponding to isoform ROR2-202 and isoforms ROR2-201/202 together can be seen. Amplicons were separated on a 2% agarose gel and visualized using Ethidium bromide staining.

Article Snippet: After blocking overnight at 4°C, immunoblots were incubated with primary anti-ROR2 antibody (1:500, ABIN2706970, Cohesion Biosciences, Aachen, Germany) and anti-β-actin (1:10,000, A5441, Sigma Aldrich, Darmstadt, Germany) at room temperature for 1 h. Incubation with the secondary antibodies (1:5000 for #1706515, 1:10,000 for #1706516, Bio-Rad, Munich, Germany) was done at room temperature for 1 h. Immunoblots were developed with Western ECL (GERPN2109, Sigma Aldrich, Darmstadt, Germany).

Techniques: Expressing, Control, Agarose Gel Electrophoresis, Staining

Journal: Frontiers in Genetics

Article Title: Interdigital Hyperplasia in Holstein Cattle Is Associated With a Missense Mutation in the Signal Peptide Region of the Tyrosine-Protein Kinase Transmembrane Receptor Gene

doi: 10.3389/fgene.2019.01157

Figure Lengend Snippet: Comparison of the receptor tyrosine kinase-like orphan receptor 2 ( ROR2 ) expression in hyperplastic interdigital skin tissue. Interdigital skin tissues were collected from 3 A_A, 3 A_T, 2 T_T cattle using fine needle biopsies (FNB). Genotypes A_A, A_T and T_T correspond to the missense variant in exon 1 (rs377953295). Two samples were taken from healthy control cattle as reference. Expression of ROR2 and internal control was done by real-time quantitative PCR. Calculation of ROR2 expression fold change in the IH affected cattle was done using the 2 -ΔΔCt method with β-actin as normalizer and the average expression of ROR2 in the healthy controls as reference . ROR2 expression fold change of the three genotypes is shown as box-and-whisker plot (whiskers indicate minima and maxima). The vertical line depicts the median. Significance was calculated using a one-tailed t-test with p < 0.05.

Article Snippet: After blocking overnight at 4°C, immunoblots were incubated with primary anti-ROR2 antibody (1:500, ABIN2706970, Cohesion Biosciences, Aachen, Germany) and anti-β-actin (1:10,000, A5441, Sigma Aldrich, Darmstadt, Germany) at room temperature for 1 h. Incubation with the secondary antibodies (1:5000 for #1706515, 1:10,000 for #1706516, Bio-Rad, Munich, Germany) was done at room temperature for 1 h. Immunoblots were developed with Western ECL (GERPN2109, Sigma Aldrich, Darmstadt, Germany).

Techniques: Comparison, Expressing, Variant Assay, Control, Real-time Polymerase Chain Reaction, Whisker Assay, One-tailed Test

Journal: Frontiers in Genetics

Article Title: Interdigital Hyperplasia in Holstein Cattle Is Associated With a Missense Mutation in the Signal Peptide Region of the Tyrosine-Protein Kinase Transmembrane Receptor Gene

doi: 10.3389/fgene.2019.01157

Figure Lengend Snippet: Quantification of the receptor tyrosine kinase-like orphan receptor 2 (ROR2) protein in hyperplastic interdigital skin tissue. Interdigital skin tissues were collected from 3 A_A, 3 A_T, 1 T_T cattle using fine needle biopsies (FNB) (see also Figure 4 ). Genotypes A_A, A_T and T_T correspond to the missense variant in exon 1 (rs377953295). Two samples were taken from healthy control cattle as reference. Protein extracts were separated on 8% Bolt Bis-Tris Plus gels and transferred to nitrocellulose membranes. Membranes were incubated with primary anti-ROR2 antibody and anti-β-actin. Blots were developed with Western ECL. Images were captured and intensity quantification was performed with ImageJ software . Significance was calculated using a one-tailed t-test with p < 0.05. As only one samples for genotype T_T were available statistical significance, average and standard deviation is not depicted. Whiskers indicate minima and maxima. The vertical line depicts the median.

Article Snippet: After blocking overnight at 4°C, immunoblots were incubated with primary anti-ROR2 antibody (1:500, ABIN2706970, Cohesion Biosciences, Aachen, Germany) and anti-β-actin (1:10,000, A5441, Sigma Aldrich, Darmstadt, Germany) at room temperature for 1 h. Incubation with the secondary antibodies (1:5000 for #1706515, 1:10,000 for #1706516, Bio-Rad, Munich, Germany) was done at room temperature for 1 h. Immunoblots were developed with Western ECL (GERPN2109, Sigma Aldrich, Darmstadt, Germany).

Techniques: Variant Assay, Control, Incubation, Western Blot, Software, One-tailed Test, Standard Deviation

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: (A) Immunohistochemical analysis of human specimens of urothelial carcinoma of the bladder for the expression of Wnt5a, ROR2, CTHRC1 and E-cadherin. Left column, tissue sections from a representative case of low grade urothelial carcinoma; the right column, tissue sections from a representative case of high grade urothelial carcinoma. The middle column represents the trend of expression for each protein in all 15 samples. The expression of Wnt5a, ROR2, and CTHRC1 increases in high grade tumors while E-cadherin shows an opposite trend. Bar=50 μm. (B) Statistical analysis was performed to investigate the correlation between tumor histological grade and immunostaining for Wnt5a, Ror2, CTHRC1, and E-cadherin. Statistical significance was tested at an alpha of 0.05. The software PASW Statistics 18 was used for data analysis (Pearson Education, New York City, NY).

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques: Immunohistochemical staining, Expressing, Immunostaining, Software

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: Wnt5a (A) , Ror2 and CTHRC1 (B) , E-cadherin and vimentin (C) , in RT4, J82 and T24 urothelial carcinoma cell lines.

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques:

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: Top row: H&E stain shows morphological differences between each cell line. Middle row: confocal microscopy images show the expression of Wnt5a (green) for each cell line. Bottom row: confocal microscopy image of the merge expression of Ror2 (green) and CTHRC1 (red) for each cell line. Although the co-expression and colocalization of Ror2 and CTHRC1 is present in all cell lines, it is clearest for RT4 and J82 cell lines.

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques: Staining, Confocal Microscopy, Expressing

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: Primer sequences used for real-time RT-PCR analyses

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques: Quantitative RT-PCR

Journal: The Journal of Experimental Medicine

Article Title: CD55 regulates self-renewal and cisplatin resistance in endometrioid tumors

doi: 10.1084/jem.20170438

Figure Lengend Snippet: CD55 localization to lipid rafts is essential for its signaling via ROR2-JNK and LCK pathways. (A) Immunofluorescent staining of cisplatin-naive non-CSCs transduced with CD55 OE, GPI-deficient transmembrane (TM)-CD55, and empty vector control. The arrowheads point to areas where CD55 is not localized to lipid rafts. (B) Graph showing the percentage of CD55–cholera toxin B colocalization. Data are representative of two independent experiments, quantifying >40 cells/group. (C) Complement-mediated cytotoxicity as assessed by the percentage BCECF dye release in A2780 non-CSCs transduced with CD55 OE, TM-CD55, and empty vector control. Data are representative of two independent experiments, and three technical replicates were used. (D) Limiting dilution analysis plots of CD55 empty vector control compared with CD55 OE and TM-CD55 constructs in cisplatin-naive non-CSCs. (E) CD55 OE cisplatin-naive non-CSCs and their empty vector controls were treated with 0–50 μM cisplatin, and percentage surviving cells was graphed. Data are representative of three independent experiments. (F and G) Immunoblots of cisplatin-naive CSCs silenced for CD55 using two shRNA constructs and a nontargeting control were probed with CD55, ROR2, pJNK (T183/Y185), JNK, pLCK (Y394), and LCK. Actin was used as a loading control. Data are representative of two independent experiments. (H and I) Cell lysates from cisplatin-naive non-CSCs transduced with CD55 and empty vector control were probed for CD55, ROR2, pJNK (T183/Y185), JNK, pLCK (Y394), and LCK. Actin was used as a loading control. Data are representative of two independent experiments. (J) Immunoblots of cisplatin-naive non-CSCs transduced with CD55, TM-CD55, and empty vector control were probed with CD55, ROR2, pLCK (Y394), and LCK. Actin was used as a loading control. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Bar, 4 µm.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against CD55 (1:1,000; Santa Cruz), CD59 (1:1,000; Abcam), CD46 (1:1,000; Santa Cruz), NANOG (1:500; Cell Signaling), SOX2 (1:500; Cell Signaling), OCT4 (1:500; Cell Signaling), ROR2 (1:1,000; BD Biosciences), pJNK (1:1,000; T183/Y185; Cell Signaling), JNK (1:1,000; Cell Signaling), pLCK (Y394; 1:1,000; BD Biosciences), LCK (1:1,000; Santa Cruz), LIME (1:1,000; Invitrogen), PAG (1:1,000; Genetex), and β-actin (1:1,000; Cell Signaling).

Techniques: Staining, Transduction, Plasmid Preparation, Flow Cytometry Complement-Mediated Cytotoxicity Assay, Construct, Western Blot, shRNA

Journal: The Journal of Experimental Medicine

Article Title: CD55 regulates self-renewal and cisplatin resistance in endometrioid tumors

doi: 10.1084/jem.20170438

Figure Lengend Snippet: LIME is necessary for intracellular CD55 signaling. (A) Immunoprecipitation (IP) experiments with CD55 antibody were performed in cisplatin-naive CSCs, and eluates were probed for lipid raft adaptor proteins LIME and PAG. (B) Cell lysates from LIME-silenced A2780 CSCs and their nontargeted (NT) controls were immunoblotted and probed with LIME, ROR2, pLCK (Y394), and LCK. Actin was used as loading control. (C) IP experiments with CD55 antibody were performed in LIME-silenced and NT control cisplatin-naive CSCs and eluates were probed for ROR2, pLCK (Y394), LCK, LIME, and CD55. (D) Immunoblots of cisplatin-naive CSCs with LIME-silenced and NT controls were immunoblotted for LIME, NANOG, SOX2, and OCT4. Actin was used as a loading control. (E) Limiting dilution analysis of LIME NT control compared with LIME sh1 and sh2 silencing constructs in cisplatin-naive CSCs. (F) LIME-silenced cisplatin-naive CSCs and their NT controls were treated with 0–50 μM cisplatin, and percentage of surviving cells is graphed. All data are representative of two or three independent experiments. **, P < 0.01; ***, P < 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against CD55 (1:1,000; Santa Cruz), CD59 (1:1,000; Abcam), CD46 (1:1,000; Santa Cruz), NANOG (1:500; Cell Signaling), SOX2 (1:500; Cell Signaling), OCT4 (1:500; Cell Signaling), ROR2 (1:1,000; BD Biosciences), pJNK (1:1,000; T183/Y185; Cell Signaling), JNK (1:1,000; Cell Signaling), pLCK (Y394; 1:1,000; BD Biosciences), LCK (1:1,000; Santa Cruz), LIME (1:1,000; Invitrogen), PAG (1:1,000; Genetex), and β-actin (1:1,000; Cell Signaling).

Techniques: Immunoprecipitation, Western Blot, Construct

Journal: The Journal of Experimental Medicine

Article Title: CD55 regulates self-renewal and cisplatin resistance in endometrioid tumors

doi: 10.1084/jem.20170438

Figure Lengend Snippet: CD55 signals via ROR2-JNK pathway to regulate self-renewal. (A) Cell lysates from cisplatin-naive CSCs and non-CSCs were immunoblotted for ROR2, pJNK (T183/Y185), and JNK. Actin was used as a loading control. (B) Immunoprecipitation (IP) analysis with CD55 antibody were performed in cisplatin-naive CSCs, and eluates were immunoblotted for ROR2. (C) Immunoblots of ROR2 silenced using two shRNA constructs and nontargeting constructs in cisplatin-naive CSCs for ROR2, pJNK (T183/Y185), JNK, NANOG, SOX2, and OCT4. Actin was used as a loading control. (D) ROR2 silenced and NT controlled A2780 CSCs analyzed by flow cytometry for GFP intensity, which indicates NANOG promoter activity. (E) Limiting dilution analysis of CD55 NT control compared with ROR2 sh1 and sh2 silencing constructs in cisplatin-naive CSCs. (F) ROR2-silenced cisplatin-naive CSCs and their NT controls were treated with 0–50 µM cisplatin, and percentage surviving cells is graphed. All data are representative of two or three independent experiments. ***, P < 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against CD55 (1:1,000; Santa Cruz), CD59 (1:1,000; Abcam), CD46 (1:1,000; Santa Cruz), NANOG (1:500; Cell Signaling), SOX2 (1:500; Cell Signaling), OCT4 (1:500; Cell Signaling), ROR2 (1:1,000; BD Biosciences), pJNK (1:1,000; T183/Y185; Cell Signaling), JNK (1:1,000; Cell Signaling), pLCK (Y394; 1:1,000; BD Biosciences), LCK (1:1,000; Santa Cruz), LIME (1:1,000; Invitrogen), PAG (1:1,000; Genetex), and β-actin (1:1,000; Cell Signaling).

Techniques: Immunoprecipitation, Western Blot, shRNA, Construct, Flow Cytometry, Activity Assay

Journal: The Journal of Experimental Medicine

Article Title: CD55 regulates self-renewal and cisplatin resistance in endometrioid tumors

doi: 10.1084/jem.20170438

Figure Lengend Snippet: CD55 regulates self-renewal and cisplatin resistance in endometrioid tumors. CD55 is glycophosphatidylinositol (GPI)–anchored to lipid rafts and via LIME binding signals intracellularly to ROR2 and LCK. ROR2 via JNK signaling regulates pluripotency gene expression, namely NANOG, SOX2, and OCT4 to maintain stemness in CSCs. In parallel, CD55 via the LCK pathway promotes the expression of DNA repair genes (including BRCA1 and MLH1) to drive cisplatin resistance.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against CD55 (1:1,000; Santa Cruz), CD59 (1:1,000; Abcam), CD46 (1:1,000; Santa Cruz), NANOG (1:500; Cell Signaling), SOX2 (1:500; Cell Signaling), OCT4 (1:500; Cell Signaling), ROR2 (1:1,000; BD Biosciences), pJNK (1:1,000; T183/Y185; Cell Signaling), JNK (1:1,000; Cell Signaling), pLCK (Y394; 1:1,000; BD Biosciences), LCK (1:1,000; Santa Cruz), LIME (1:1,000; Invitrogen), PAG (1:1,000; Genetex), and β-actin (1:1,000; Cell Signaling).

Techniques: Binding Assay, Expressing

Journal: Scientific Reports

Article Title: High ROR2 expression in tumor cells and stroma is correlated with poor prognosis in pancreatic ductal adenocarcinoma

doi: 10.1038/srep12991

Figure Lengend Snippet: Two PDAC cell lines (BXPC3, PNAC-1) have higher ROR2 expression than the benign pancreatic ductal cell line (HPDE6C7). The mean expression level of ROR2 mRNA in cancerous tissue was higher on average in the tumor tissues than in corresponding non-malignant tissues.

Article Snippet: TMA slides were separately stained using polyclonal rabbit anti-ROR2 antibody (LifeSpan BioSciences Inc., Seattle, WA, USA).

Techniques: Expressing

Journal: Scientific Reports

Article Title: High ROR2 expression in tumor cells and stroma is correlated with poor prognosis in pancreatic ductal adenocarcinoma

doi: 10.1038/srep12991

Figure Lengend Snippet: ( a1 , a2 ) Positive tumor cytoplasmic (red arrow) and negative stromal (blue arrow) immunohistochemical staining of ROR2 in PDAC samples. ( b1 , b2 ) Strong tumor (red arrow) and stromal (green arrow) immunohistochemical staining of ROR2 in PDAC samples. ( c1 , c2 ) Negative IHC of ROR2 in tumor cells (black arrow) with strong positive stromal (green arrow) staining in PDAC tissue samples. ( d1 , d2 ) There was negative immunohistochemical staining of ROR2 in cancer cells (black arrow) and stromal cells (blue arrow). ( e1 , e2 ) Negative staining of ROR2 in epithelial cells (black arrow) in benign pancreatic disease tissues. ( f1 , f2 ) Negative immunohistochemical staining of ROR2 in benign pancreatic ductal cells (black arrow). Original magnification ×40 (bar = 500 μm) in ( a1 , b1 , c1 , d1 , e1 and f1 ); ×400 (bar = 50 μm) in ( a2 , b2 , c2 , d2 , e2 and f2 ).

Article Snippet: TMA slides were separately stained using polyclonal rabbit anti-ROR2 antibody (LifeSpan BioSciences Inc., Seattle, WA, USA).

Techniques: Immunohistochemical staining, Staining, Negative Staining

Journal: Scientific Reports

Article Title: High ROR2 expression in tumor cells and stroma is correlated with poor prognosis in pancreatic ductal adenocarcinoma

doi: 10.1038/srep12991

Figure Lengend Snippet: Association of ROR2 expression with clinical attributes of pancreatic ductal adenocarcinoma.

Article Snippet: TMA slides were separately stained using polyclonal rabbit anti-ROR2 antibody (LifeSpan BioSciences Inc., Seattle, WA, USA).

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: High ROR2 expression in tumor cells and stroma is correlated with poor prognosis in pancreatic ductal adenocarcinoma

doi: 10.1038/srep12991

Figure Lengend Snippet: Univariate and multivariate analysis of prognostic factors for 5-year survival in pancreatic cancer.

Article Snippet: TMA slides were separately stained using polyclonal rabbit anti-ROR2 antibody (LifeSpan BioSciences Inc., Seattle, WA, USA).

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: High ROR2 expression in tumor cells and stroma is correlated with poor prognosis in pancreatic ductal adenocarcinoma

doi: 10.1038/srep12991

Figure Lengend Snippet: ( a ) Overall survival rate in patients with high cytoplasmic ROR2 expression (green line) was significantly lower than in patients with low or no cytoplasmic ROR2 expression (blue line). ( b ) Overall survival rate in patients with high stromal ROR2 expression (green line) was significantly lower than in patients with low or no stromal ROR2 expression (blue line).

Article Snippet: TMA slides were separately stained using polyclonal rabbit anti-ROR2 antibody (LifeSpan BioSciences Inc., Seattle, WA, USA).

Techniques: Expressing