Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: KTN0158, a humanized anti-KIT monoclonal antibody, demonstrates biologic activity against both normal and malignant canine mast cells

doi: 10.1158/1078-0432.CCR-16-2152

Figure Lengend Snippet: (A) KTN0158 inhibited KIT activation and downstream signaling in M-07e cells. Dose-dependent inhibition of KIT, ERK1/2 and AKT activation were observed flowing KTN0158 treatment. KTN0062C is an isotype control antibody. (B) KTN0158 was a more potent inhibitor of SCF-dependent proliferation in M-07e cells compared to imatinib and nilotinib. Dashed lines represent proliferation observed in controls grown in the presence (+SCF) or absence (-SCF) of SCF. Representative data are shown for six independent experiments. Analyses were performed in duplicate (KTN0158) or triplicate (all others) and data are plotted as means ± standard error of the mean (SEM). (C) KTN0158 was a more potent inhibitor of SCF-induced KIT phosphorylation in primary human mast cells in vitro than imatinib. Representative data are shown for four independent mast cell preparations and runs. Analyses were performed in duplicate and data are plotted as means ± SEM. (D) KTN0158 was a potent inhibitor of SCF enhancement of mast cell degranulation. In both (C) and (D), the isotype control is KTN0062C and analyses were performed in duplicate with data plotted as means ± SEM.

Article Snippet: The human acute megakaryoblastic leukemia cell line M-07e ( 19 ) was obtained from the Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) GmbH (Braunschweig, Germany) and used within 6 months of receipt.

Techniques: Activation Assay, Inhibition, Control, Phospho-proteomics, In Vitro

Journal: Molecular and Clinical Oncology

Article Title: Analysis of genes encoding epigenetic regulators in myeloproliferative neoplasms: Coexistence of a novel SETBP1 mutation in a patient with a p.V617F JAK2 positive myelofibrosis

doi: 10.3892/mco.2019.1840

Figure Lengend Snippet: Human cancer cell lines tested.

Article Snippet: Informed consent was obtained from all patients and procedures were approved by the Ethical Committee on Clinical Research of University of Navarra. table ft1 table-wrap mode="anchored" t5 Table II. caption a7 Cell line name Repository (number) Origin A-549 DSMZ (ACC-107) Lung carcinoma BK-006 ECACC p.V617F JAK2 positive PV BK-013 ECACC (98100924) p.V617F JAK2 negative ET BK015 ECACC (99092421) p.V617F JAK2 negative ET DAUDI DSMZ (ACC-78) Burkitt lymphoma EOL-1 DSMZ (ACC-386) Acute myeloid eosinophilic leukemia F-36P DSMZ (ACC-543) Acute myeloid leukemia secondary to myelodysplastic syndrome (MDS) HCC-1937 DSMZ (ACC-513) Breast carcinoma HEL DSMZ ACC-11 Erythroleukemia HL60 DSMZ ACC-3 Acute myeloid leukemia (AML) HU3 Dr Morgan, USA Acute megakaryoblastic leukemia K-562 DSMZ ACC-10 BCR-ABL1 positive chronic myeloid leukemia (CML) in blast crisis KARPAS-299 DSMZ ACC-31 T cell lymphoma KARPAS-422 DSMZ ACC-32 B cell lymphoma M-07e DSMZ ACC-104 Acute megakaryoblastic leukemia MG-63 ATCC: CRL-1427 Osteosarcoma MOLM-13 DSMZ ACC-554 Acute myeloid leukemia (AML) MOLT-16 DSMZ ACC-29 T cell acute lymphoid leukemia (T-ALL) MOLT-4 DSMZ ACC-362 Acute lymphoid leukemia (ALL) MV-411 DSMZ ACC-102 Acute monoblastic/monocytic leukemia RAJI DSMZ ACC-319 Burkitt lymphoma REH DSMZ ACC-22 B-ALL (t(12;21) (p13;q22) (fusion ETV6-RUNX1 )) SET2 DSMZ ACC-608 p.V617F JAK2 positive ET TF-1 DSMZ ACC-334 Erythroleukemia UKE-1 Dr W. Fiedelr, Hamburg, Germany p.V617F JAK2 positive ET transformed to AML Open in a separate window ECACC, European Collection of Cell Culture; DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen.

Techniques: Transformation Assay

Journal: Molecular and Clinical Oncology

Article Title: Analysis of genes encoding epigenetic regulators in myeloproliferative neoplasms: Coexistence of a novel SETBP1 mutation in a patient with a p.V617F JAK2 positive myelofibrosis

doi: 10.3892/mco.2019.1840

Figure Lengend Snippet: Human cancer cell lines tested.

Article Snippet: Informed consent was obtained from all patients and procedures were approved by the Ethical Committee on Clinical Research of University of Navarra. table ft1 table-wrap mode="anchored" t5 Table II. caption a7 Cell line name Repository (number) Origin A-549 DSMZ (ACC-107) Lung carcinoma BK-006 ECACC p.V617F JAK2 positive PV BK-013 ECACC (98100924) p.V617F JAK2 negative ET BK015 ECACC (99092421) p.V617F JAK2 negative ET DAUDI DSMZ (ACC-78) Burkitt lymphoma EOL-1 DSMZ (ACC-386) Acute myeloid eosinophilic leukemia F-36P DSMZ (ACC-543) Acute myeloid leukemia secondary to myelodysplastic syndrome (MDS) HCC-1937 DSMZ (ACC-513) Breast carcinoma HEL DSMZ ACC-11 Erythroleukemia HL60 DSMZ ACC-3 Acute myeloid leukemia (AML) HU3 Dr Morgan, USA Acute megakaryoblastic leukemia K-562 DSMZ ACC-10 BCR-ABL1 positive chronic myeloid leukemia (CML) in blast crisis KARPAS-299 DSMZ ACC-31 T cell lymphoma KARPAS-422 DSMZ ACC-32 B cell lymphoma M-07e DSMZ ACC-104 Acute megakaryoblastic leukemia MG-63 ATCC: CRL-1427 Osteosarcoma MOLM-13 DSMZ ACC-554 Acute myeloid leukemia (AML) MOLT-16 DSMZ ACC-29 T cell acute lymphoid leukemia (T-ALL) MOLT-4 DSMZ ACC-362 Acute lymphoid leukemia (ALL) MV-411 DSMZ ACC-102 Acute monoblastic/monocytic leukemia RAJI DSMZ ACC-319 Burkitt lymphoma REH DSMZ ACC-22 B-ALL (t(12;21) (p13;q22) (fusion ETV6-RUNX1 )) SET2 DSMZ ACC-608 p.V617F JAK2 positive ET TF-1 DSMZ ACC-334 Erythroleukemia UKE-1 Dr W. Fiedelr, Hamburg, Germany p.V617F JAK2 positive ET transformed to AML Open in a separate window ECACC, European Collection of Cell Culture; DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen.

Techniques: Transformation Assay

Journal: Journal of Thrombosis and Haemostasis

Article Title: Increased expression of HIF2α during iron deficiency–associated megakaryocytic differentiation

doi: 10.1111/jth.12884

Figure Lengend Snippet: Regulation of genes in the hypoxia‐inducible factor (HIF) and vascular endothelial growth factor (VEGF) pathways is retained in subsequent experiments with iron deficiency (ID). Verification of selected identified regulated genes via real time quantitative PCR (RT‐PCR) in human erythroleukemia (HEL) (A) and megakaryoblastic leukemia (CMK) (B) after 3 days and megakaryocytes (MEGs) (C) and CD61‐negative cells (D) after 5 days of ID and iron‐replete condition (IR). Graphs depict the relative mRNA quantity (RQ) of selected genes in cells cultured under ID as compared to IR. All results are normalized to HPRT1 .

Article Snippet: Human erythroleukemia (HEL) and megakaryoblastic leukemia (CMK) human cell lines (DSMZ, Braunschweig, Germany) were maintained in RPMI (Gibco/Life Technologies, Lofer, Austria) and adapted to serum‐free and iron‐replete (IR) medium (Panserin 401; PanBiotech, Aidenbach, Germany) before experiments.

Techniques: Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Cell Culture

Journal: The Journal of biological chemistry

Article Title: Purification of storage granule protein-23. A novel protein identified by phage display technology and interaction with type I plasminogen activator inhibitor.

doi: 10.1074/jbc.271.47.30126

Figure Lengend Snippet: FIG. 2. Application of clone A-61 for the analysis of AtT-20 RNA and protein. A, Northern blotting analysis of AtT-20 and DAMI cell mRNA with clone A-61. Total RNA was isolated from AtT-20 (lane 1) and DAMI (lane 2) cells, subjected to agarose electrophoresis (10 mg/ lane), transferred to nitrocellulose, and probed with a 32P-labeled A-61 probe. The positions of 28 and 18 S ribosomal RNA are indicated. B, prokaryotic expression of pTrcHis/A-61. pTrcHis/A-61-transformed (lanes 1 and 3) or pTrcHis-transformed (lanes 2 and 4) E. coli (1 liter of culture/preparation) were induced with isopropyl-b-D-thiogalactopy- ranoside for 4 h, and the bacteria preparations were lysed and fraction- ated on nickel-chelate columns as described under “Materials and Methods.” The material eluting between 0.25 and 0.4 M imidazole was pooled, subjected to SDS-PAGE (50 mg/lane), and analyzed by silver staining (lanes 1 and 2) or by immunoblotting employing anti-decapep- tide as the primary antibody (lanes 3 and 4). C, identification of an immunologically related protein in AtT-20 cells with anti-A-61. AtT-20 cells (lanes 1, 3, 5, and 7; 105 cells/lane) and purified A-61 (lanes 2, 4, 6, and 8; 500 ng/lane) were subjected to SDS-PAGE under reducing (lanes 1–4) or nonreducing (lanes 5–8) conditions, transferred to nitrocellu- lose, and analyzed by immunoblotting with antibodies directed against A-61 (lanes 1, 2, 5, and 6) or normal IgG (lanes 3, 4, 7, and 8).

Article Snippet: Growth of Cultured Cells—AtT-20 cells, DAMI cells (a cell line established from an individual with megakaryoblastic leukemia (39)), and a transformed human fibroblast cell line (SV40 WI38 VA13 2RA) were obtained from the American Tissue Culture Collection (Rockville, MD).

Techniques: Northern Blot, Isolation, Electrophoresis, Labeling, Expressing, Transformation Assay, Bacteria, SDS Page, Silver Staining, Western Blot, Purification