Journal: Oncology Reports

Article Title: Retinoic acid induces differentiation in neuroblastoma via ROR1 by modulating retinoic acid response elements

doi: 10.3892/or.2020.7681

Figure Lengend Snippet: RA promotes NB differentiation via ROR1. (A) Immunoblotting revealed the changes in ROR1 and synaptophysin levels under normal differentiation with RA. (B) Densitometry of immunoblots from A, baseline-corrected to GAPDH (ANOVA with Tukey's multiple comparison to all conditions, n=3). **P<0.005, ***P<.0005, ****P<0.0001. (C) Phase contrast images of SK-N-SH neuroblastoma cells undergoing RA or DMSO treatment over 96 h under shRNA test Lipofectamine transfection, knocking down ROR1 expression. Yellow arrows indicate marked neurite extensions that suggest development. (D) Visualization of the average neurite length over time following knockdown of ROR1 (n=3). (E) Immunoblots revealed the changes in ROR1 and synaptophysin levels after Lipofectamine transfection, knocking down ROR1 expression. (F) Densitometry of immunoblots from E, baseline-corrected to GAPDH (ANOVA with Tukey's multiple comparison test to compare each condition with every other condition in the experiment, n=3). ****P<0.0001. (G) Immunofluorescence revealed the effects of RA on synaptophysin (green) levels with or without shRNA knockdown of ROR1 (red). DAPI (blue) highlights the nucleus of the neurons. Yellow arrows indicate marked neurite extensions. RA, retinoic acid; NB, neuroblastoma; ROR1, receptor tyrosine kinase-like orphan receptor 1; NT, not treated.

Article Snippet: The primary antibodies used were as follows and diluted 1:1,000 unless specifically specified differently under manufacturer recommendations: ROR1 (D6T8C; product no. 16540; Cell Signaling Technology, Inc.), GAPDH (D16H11; product no. 5174; Cell Signaling Technology, Inc.), p-AKT-ser473 (product no. 9271; Cell Signaling Technology, Inc.), AKT (C67E7; product no. 4691; Cell Signaling Technology, Inc.), p-GSK3β-Ser9 (D17D2; product no. 8566; Cell Signaling Technology, Inc.), and GSK3β (D75D3; product no. 5676; Cell Signaling Technology, Inc.), β-catenin (cat. no. MA1-301; Invitrogen; Thermo Fisher Scientific, Inc.), p-β-catenin-Ser33/37 (product no. 2009; Cell Signaling Technology, Inc.), stabilized-β-catenin (product no. 9562; Cell Signaling Technology, Inc.), Wnt-5a (cat. no. NBP2-24752SS; Novus Biologicals, Ltd.), Synaptophysin (cat. no. 100298-T40; Sino Biological, Inc.), β-tubulin (product code ab21057; Abcam) and Lamin B1 (product code ab65986; Abcam).

Techniques: Western Blot, shRNA, Transfection, Expressing, Immunofluorescence

Journal: Oncology Reports

Article Title: Retinoic acid induces differentiation in neuroblastoma via ROR1 by modulating retinoic acid response elements

doi: 10.3892/or.2020.7681

Figure Lengend Snippet: Effects of RA on ROR1 and differentiation is RAR-dependent. (A) Light-microscopic images (×100) after various treatments modulating the degree of ROR1 in the cell at 0 and 96 h. Yellow arrows indicate marked neurite lengths and neurite clusters that suggest progression of development. (B) Visualization of average neurite length after ROR1-modulating treatments over 96 h (n=3). *P<0.05. (C) Immunoblots of synaptophysin and ROR1 after ROR1-modulating treatments. (D) Densitometric analysis of immunoblots in C assessing ROR1 and synaptophysin levels (ANOVA with Tukey's multiple comparison test to compare each condition with every other condition in the experiment, n=3). **P<0.005, ****P<0.0001. RA, retinoic acid; ROR1, receptor tyrosine kinase-like orphan receptor 1; RAR, RA receptor.

Article Snippet: The primary antibodies used were as follows and diluted 1:1,000 unless specifically specified differently under manufacturer recommendations: ROR1 (D6T8C; product no. 16540; Cell Signaling Technology, Inc.), GAPDH (D16H11; product no. 5174; Cell Signaling Technology, Inc.), p-AKT-ser473 (product no. 9271; Cell Signaling Technology, Inc.), AKT (C67E7; product no. 4691; Cell Signaling Technology, Inc.), p-GSK3β-Ser9 (D17D2; product no. 8566; Cell Signaling Technology, Inc.), and GSK3β (D75D3; product no. 5676; Cell Signaling Technology, Inc.), β-catenin (cat. no. MA1-301; Invitrogen; Thermo Fisher Scientific, Inc.), p-β-catenin-Ser33/37 (product no. 2009; Cell Signaling Technology, Inc.), stabilized-β-catenin (product no. 9562; Cell Signaling Technology, Inc.), Wnt-5a (cat. no. NBP2-24752SS; Novus Biologicals, Ltd.), Synaptophysin (cat. no. 100298-T40; Sino Biological, Inc.), β-tubulin (product code ab21057; Abcam) and Lamin B1 (product code ab65986; Abcam).

Techniques: Western Blot

Journal: Oncology Reports

Article Title: Retinoic acid induces differentiation in neuroblastoma via ROR1 by modulating retinoic acid response elements

doi: 10.3892/or.2020.7681

Figure Lengend Snippet: RA pathway model. The proposed proliferation and differentiation modulation pathway of RA. RA binds to RAR in the cytoplasm and migrates into the nucleus to act on RARE regions. Normal transcription and translation result in increased ROR1 and Wnt5a synthesis and the ROR1 signaling pathway cascades to synaptophysin as a node of differentiation as well as β-catenin-related growth and proliferation factors. The image was created using BioRender. RA, retinoic acid; ROR1, receptor tyrosine kinase-like orphan receptor 1; RAR, RA receptor; RARE, RA response element.

Article Snippet: The primary antibodies used were as follows and diluted 1:1,000 unless specifically specified differently under manufacturer recommendations: ROR1 (D6T8C; product no. 16540; Cell Signaling Technology, Inc.), GAPDH (D16H11; product no. 5174; Cell Signaling Technology, Inc.), p-AKT-ser473 (product no. 9271; Cell Signaling Technology, Inc.), AKT (C67E7; product no. 4691; Cell Signaling Technology, Inc.), p-GSK3β-Ser9 (D17D2; product no. 8566; Cell Signaling Technology, Inc.), and GSK3β (D75D3; product no. 5676; Cell Signaling Technology, Inc.), β-catenin (cat. no. MA1-301; Invitrogen; Thermo Fisher Scientific, Inc.), p-β-catenin-Ser33/37 (product no. 2009; Cell Signaling Technology, Inc.), stabilized-β-catenin (product no. 9562; Cell Signaling Technology, Inc.), Wnt-5a (cat. no. NBP2-24752SS; Novus Biologicals, Ltd.), Synaptophysin (cat. no. 100298-T40; Sino Biological, Inc.), β-tubulin (product code ab21057; Abcam) and Lamin B1 (product code ab65986; Abcam).

Techniques:

Journal: Cancer cell

Article Title: Binary pan-cancer classes with distinct vulnerabilities defined by pro- or anti-cancer YAP/TEAD activity

doi: 10.1016/j.ccell.2021.06.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal anti-Synaptophysin (clone MRQ-40) , Origene , Cat# TA327740.

Techniques: Virus, Plasmid Preparation, Derivative Assay, Recombinant, Electron Microscopy, Staining, SYBR Green Assay, Reverse Transcription, Sample Prep, Microarray, Knockdown, Gene Expression, CRISPR, Software, Flow Cytometry

Journal: OncoTargets and therapy

Article Title: An isolated orbital mass with proptosis and diplopia: an uncommon metastasis from intracranial olfactory neuroblastoma in a chinese patient

doi: 10.2147/OTT.SI09929

Figure Lengend Snippet: Immunohistochemical staining of the mass. Notes: The tumor cells were positive for ( A ) SYN; original magnification: ×400; a reaction in the cytoplasm of the neoplastic cells; ( B ) CGA; original magnification: ×400; a reaction in the cytoplasm of the neoplastic cells; (C) CD56 (cluster of differentiation 56); original magnification: ×400; a strong and heavy membrane-type staining; ( D ) CK; original magnification: ×400; a reaction in the cytoplasm of the neoplastic cells; ( E ) VIM; original magnification: x400; a reaction in the cytoplasm of the neoplastic cells; ( F ) S-100 protein; original magnification: ×400; a sustentacular reaction. The tumor cells were negative for EMA and CD99 (data not shown). Abbreviations: CD, cluster of differentiation; CGA, chromogranin; CK, cytokeratin; EMA, epithelial membrane antigen; SYN, synaptophysin; VIM, vimentin.

Article Snippet: Immunohistochemical analyses for synaptophysin (SYN) (rabbit monoclonal antibody, OriGene, Rockville, MD, USA), chromogranin A (CGA; mouse monoclonal antibody; OriGene), cluster of differentiation (CD56; mouse monoclonal antibody; OriGene), cytokeratin (CK; mouse monoclonal antibody; OriGene), vimentin (VIM; mouse monoclonal antibody; OriGene), S-100 protein (mouse monoclonal antibody, OriGene), epithelial membrane antigen (EMA; mouse monoclonal antibody; OriGene), and cluster of differentiation 99 (CD99; mouse monoclonal antibody; OriGene) were performed on the paraffin sections.

Techniques: Immunohistochemical staining, Staining, Membrane

Journal: OncoTargets and therapy

Article Title: An isolated orbital mass with proptosis and diplopia: an uncommon metastasis from intracranial olfactory neuroblastoma in a chinese patient

doi: 10.2147/OTT.SI09929

Figure Lengend Snippet: Selected immunohistochemical results for malignant small round cell tumors

Article Snippet: Immunohistochemical analyses for synaptophysin (SYN) (rabbit monoclonal antibody, OriGene, Rockville, MD, USA), chromogranin A (CGA; mouse monoclonal antibody; OriGene), cluster of differentiation (CD56; mouse monoclonal antibody; OriGene), cytokeratin (CK; mouse monoclonal antibody; OriGene), vimentin (VIM; mouse monoclonal antibody; OriGene), S-100 protein (mouse monoclonal antibody, OriGene), epithelial membrane antigen (EMA; mouse monoclonal antibody; OriGene), and cluster of differentiation 99 (CD99; mouse monoclonal antibody; OriGene) were performed on the paraffin sections.

Techniques: Immunohistochemical staining