A5060895990- Ovarian cancer diagnosis and treatment
- Sarcoma Diagnosis and Treatment
- Cancer, Hypoxia, and Metabolism
- Renal and related cancers
- PI3K/AKT/mTOR signaling in cancer
- RNA modifications and cancer
- Cancer therapeutics and mechanisms
- RNA Research and Splicing
- Protein Degradation and Inhibitors
Cincinnati Children's Hospital Medical Center
2022-2023
University of Cincinnati Medical Center
2022-2023
Targeted cancer therapeutics have not significantly benefited patients with Ewing sarcoma metastatic or relapsed disease. Understanding the molecular underpinnings of drug resistance can lead to biomarker-driven treatment selection.Receptor tyrosine kinase (RTK) pathway activation was analyzed in tumor cells derived from a panel tumors, including primary and tumors same patient. Phospho-RTK arrays, Western blots, IHC were used. Protein localization levels key markers determined using...
<p>Immunofluorescence images of CCH2 and NCH6 cells stained with antibodies towards IGF1R (red) PCNA (green).</p>
<p>Western blots and quantifications</p>
<p>Reagents, instruments, tumor cell culture details</p>
<p>Cell cycle analyses A. Dansylcadaverine-mediated nIGF1R depletion arrested CCH1 cells at the G1-phase and reduced percentage of in S-phase. B. Western blots show that Cyclin A2 D1 levels were high CCH5 with relative to NCH6 mIGF1R. C. Cell shows higher percentages S-phase than CCH2 cells. D. divided faster, had shorter duration E. EdU/BrdU double labelling-based analysis was carried out. Top panel showed EdU-AF55+ BrdU-FITC+ signals comparison unstained F. Percentages EdU+BrdU+...
<p>Flow cytometry analysis for propidium-iodide positive cells with the indicated treatments (48 hours). The percentage of PI-positive are shown in each panel.</p>
<p>A. Bright field images of CCH1 and CCH2 cells B. stained with antibody towards CD99 (green) DAPI (blue) C. Western blots on cell lysates probed the indicated antibodies, confirming differences in EphA2 phosphorylation between CCH2. D. Immunofluorescence imaging IGF1R-stained starved, treated IGF1 or linsitinib for 2 hours. Individual channels are shown below merged images. Nuclear IGF1R does not change localization either treatment. treatment induces slight internalization this is...
<p>Characterization of Ewing sarcoma cell lines A673, RD-ES and SK-ESA.Western blot analysis the IGF1R signaling pathway B. Western markers associated with Replication Stress Response C. RPA phosphorylation apoptosis measured by cleaved caspase 3 (CC3 levels).Cells stained antibody towards (red) DAPI (blue) shows membranous staining in SK-ES cells, very low levels A673 cells.</p>
<div>AbstractPurpose:<p>Targeted cancer therapeutics have not significantly benefited patients with Ewing sarcoma metastatic or relapsed disease. Understanding the molecular underpinnings of drug resistance can lead to biomarker-driven treatment selection.</p>Experimental Design:<p>Receptor tyrosine kinase (RTK) pathway activation was analyzed in tumor cells derived from a panel tumors, including primary and tumors same patient. Phospho-RTK arrays, Western blots, IHC...
<div>AbstractPurpose:<p>Targeted cancer therapeutics have not significantly benefited patients with Ewing sarcoma metastatic or relapsed disease. Understanding the molecular underpinnings of drug resistance can lead to biomarker-driven treatment selection.</p>Experimental Design:<p>Receptor tyrosine kinase (RTK) pathway activation was analyzed in tumor cells derived from a panel tumors, including primary and tumors same patient. Phospho-RTK arrays, Western blots, IHC...
<p>Immunofluorescence images of CCH2 and NCH6 cells stained with antibodies towards IGF1R (red) PCNA (green).</p>
<p>Patient tumor information</p>
<p>A. Representative images for DNA fiber experiments conducted on CCH1 and CCH2 cells. In each image asterisk (*) indicates a normal fork (red-green), “N” new (red only), “S” stalled (green only).B. Brightfield CD99-stained (green) of patient-derived cells used in the studies reported here.</p>
<p>Western blots and quantifications</p>
<p>Cell cycle analyses A. Dansylcadaverine-mediated nIGF1R depletion arrested CCH1 cells at the G1-phase and reduced percentage of in S-phase. B. Western blots show that Cyclin A2 D1 levels were high CCH5 with relative to NCH6 mIGF1R. C. Cell shows higher percentages S-phase than CCH2 cells. D. divided faster, had shorter duration E. EdU/BrdU double labelling-based analysis was carried out. Top panel showed EdU-AF55+ BrdU-FITC+ signals comparison unstained F. Percentages EdU+BrdU+...
<p>A. Bright field images of CCH1 and CCH2 cells B. stained with antibody towards CD99 (green) DAPI (blue) C. Western blots on cell lysates probed the indicated antibodies, confirming differences in EphA2 phosphorylation between CCH2. D. Immunofluorescence imaging IGF1R-stained starved, treated IGF1 or linsitinib for 2 hours. Individual channels are shown below merged images. Nuclear IGF1R does not change localization either treatment. treatment induces slight internalization this is...
<p>A. Representative images for DNA fiber experiments conducted on CCH1 and CCH2 cells. In each image asterisk (*) indicates a normal fork (red-green), “N” new (red only), “S” stalled (green only).B. Brightfield CD99-stained (green) of patient-derived cells used in the studies reported here.</p>
<p>Characterization of Ewing sarcoma cell lines A673, RD-ES and SK-ESA.Western blot analysis the IGF1R signaling pathway B. Western markers associated with Replication Stress Response C. RPA phosphorylation apoptosis measured by cleaved caspase 3 (CC3 levels).Cells stained antibody towards (red) DAPI (blue) shows membranous staining in SK-ES cells, very low levels A673 cells.</p>
<p>Flow cytometry analysis for propidium-iodide positive cells with the indicated treatments (48 hours). The percentage of PI-positive are shown in each panel.</p>
<p>Reagents, instruments, tumor cell culture details</p>
<p>Patient tumor information</p>