Journal: Pharmaceutics

Article Title: A Small Molecule Targeting the Intracellular Tyrosine Kinase Domain of ROR1 (KAN0441571C) Induced Significant Apoptosis of Non-Small Cell Lung Cancer (NSCLC) Cells.

doi: 10.3390/pharmaceutics15041148

Figure Lengend Snippet: Figure 1. (A) Surface staining for ROR1 (flow cytometry) (red line: isotype control; blue line: anti- ROR1 antibody); (B) Western blot for ROR1 and pROR1 expression (130 KDa) in 5 lung cancer cell lines. Actin was used as internal control.

Article Snippet: The following primary antibodies were used for protein staining: phospho (p) ROR1 (against amino acid residues Ser 652, Tyr 641, 646) [46], anti-ROR1 antibody (R&D Systems, Minneapolis, MN, USA), anti-EGFR and anti-pEGFR (Y1173) antibodies, anti-SRC and anti-p-SRC (Tyr 416) antibodies, anti-AKT and anti-p-AKT (Ser 473) antibodies, anti-mTOR and anti-p-mTOR (Ser 2448) antibodies, anti-cAMP response element-binding protein (CREB) and anti-p-CREB (Ser 133) antibodies, anti-cleaved poly ADP ribose polymerase (PARP) antibody, anti-B-cell lymphoma (BCL)-2 antibody, anti-BAX antibody, anti-myeloid cell leukemia (MCL)-1 antibody, anti-cleaved caspases 3, 8, 9 antibodies (Cell Signaling Technology, Danvers, MA, USA), anti-phosphoinositide 3-kinase (PI3K) p110δ and anti-p-PI3Kp110δ (Tyr 485) antibodies (Santa Cruz), and antiβ-actin antibody (Sigma-Aldrich).

Techniques: Staining, Cytometry, Control, Western Blot, Expressing

Journal: Molecular Oncology

Article Title: Receptor tyrosine kinase gene expression profiles of Ewing sarcomas reveal ROR1 as a potential therapeutic target in metastatic disease

doi: 10.1016/j.molonc.2015.12.009

Figure Lengend Snippet: ROR1 expression in Ewing sarcoma (A) Upper panels: ROR1 gene expression. qPCR DDCt‐method relative to GAPDH and to 697 ALL positive control cells. Graphs and error bars show mean and standard deviation of at least three independent experiments. ...

Article Snippet: Flow cytometry Cells were stained with 0.4 μg of ROR1 polyclonal antibody (R&D Systems, Minneapolis, MN) or IgG isotype control (Jackson Immunoresearch, Baltimore, PA) for 30 min at 4 °C.

Techniques: Expressing, Gene Expression, Positive Control, Standard Deviation

Journal: Molecular Oncology

Article Title: Receptor tyrosine kinase gene expression profiles of Ewing sarcomas reveal ROR1 as a potential therapeutic target in metastatic disease

doi: 10.1016/j.molonc.2015.12.009

Figure Lengend Snippet: Functional effects of ROR1 silencing A673 and 5838 cells were transfected with siRNA targeting ROR1 (siROR1) or non‐silencing control (siCTRL). (A) Left panel: Viable cells were counted by trypan blue exclusion 96 h after transfection. ...

Article Snippet: Flow cytometry Cells were stained with 0.4 μg of ROR1 polyclonal antibody (R&D Systems, Minneapolis, MN) or IgG isotype control (Jackson Immunoresearch, Baltimore, PA) for 30 min at 4 °C.

Techniques: Functional Assay, Transfection, Control

Journal: Molecular Oncology

Article Title: Receptor tyrosine kinase gene expression profiles of Ewing sarcomas reveal ROR1 as a potential therapeutic target in metastatic disease

doi: 10.1016/j.molonc.2015.12.009

Figure Lengend Snippet: Wnt5a is a potential ROR1 ligand (A) Correlation plot of ROR1 and Wnt5a gene expressions. Wnt5a gene expression was analyzed in cell lines and 12 Ewing sarcoma specimens by qPCR and ΔΔCt‐method compared to GAPDH and MSC cultures ...

Article Snippet: Flow cytometry Cells were stained with 0.4 μg of ROR1 polyclonal antibody (R&D Systems, Minneapolis, MN) or IgG isotype control (Jackson Immunoresearch, Baltimore, PA) for 30 min at 4 °C.

Techniques: Gene Expression

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: Known chemical structures of ROR1 inhibitors.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques:

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: Structure‐based drug design approach leading to the para‐ phenylenediamine derivatives acting as ROR1 inhibitors.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques:

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: CETSA identifies ROR1 protein in the binding with 17 in JeKo‐1 cells. A) CETSA western blots and B) corresponding quantitative analysis of band intensity. 17 group was treated with 17 (40 μM) for 2 h and then subjected to 5 min incubation at the indicated temperature (45–58 °C). Ctrl group was treated with vehicle (0.1% DMSO) only in the same conditions. ROR1 level at 25 °C was set at 100%. Densitometry‐based quantification of western blotting signals was calculated by first normalizing to α ‐tubulin levels in individual samples. Statistical analyses were performed comparing the compound 17 group to the Ctrl group across different temperature points. Results are shown as mean ± SD from three independent experiments. ** and *** denote respectively p < 0.01 and p < 0.001 versus Ctrl.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques: Binding Assay, Western Blot, Incubation

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: Molecular effects of 17 on ROR1 pathways in JeKo‐1 cells. For each western blotting analysis, three independent experiments were performed, and the SD is expressed by error bars. The relative fold change versus untreated cells, set as 1, is shown in the graph. *, **, and *** denote respectively p < 0.05, p < 0.01, and p < 0.001 versus Ctrl.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques: Western Blot