Additional file 1 of Therapeutic activation of G protein-coupled estrogen receptor 1 in Waldenström Macroglobulinemia
Additional file 1: Figure S1. Analysis of GPER1 mRNA in public datasets GSE9656 (A) and GSE61597 (B). GSE9656: we analyzed GPER1 mRNA in CD19-selected peripheral blood B cells (pBCs; n = 7), bone marrow B cells from WM (WM-BCs; n = 12), bone marrow plasma cells from healthy donors (BM-PCs; n = 10),...
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| creator | Morelli, Eugenio Hunter, Zachary R. Fulciniti, Mariateresa Gullà, Annamaria Perrotta, Ida Daniela Zuccalà, Valeria Federico, Cinzia Juli, Giada Manzoni, Martina Ronchetti, Domenica Romeo, Enrica Gallo Cantafio, Maria Eugenia Soncini, Debora Maltese, Lorenza Rossi, Marco Roccaro, Aldo M. Cea, Michele Tassone, Pierfrancesco Neri, Antonino Treon, Steven C. Munshi, Nikhil C. Viglietto, Giuseppe Amodio, Nicola |
| description | Additional file 1: Figure S1. Analysis of GPER1 mRNA in public datasets GSE9656 (A) and GSE61597 (B). GSE9656: we analyzed GPER1 mRNA in CD19-selected peripheral blood B cells (pBCs; n = 7), bone marrow B cells from WM (WM-BCs; n = 12), bone marrow plasma cells from healthy donors (BM-PCs; n = 10), and WM plasma cells (WM-PCs, n = 9). GSE61597: we analyzed GPER1 mRNA in normal bone marrow CD25+ (n = 7) and CD25– (n = 9) B cells, clonal B cells from newly diagnosed patients with IgM MGUS (n=22), smoldering (n = 17), and symptomatic WM (n = 10). Figure S2. A. qRT-PCR analysis of GPER1 mRNA in MCF7 breast cancer cell line (positive control), MWCL-1 and BCWM-1 WM cell lines, and CD19+ primary cells from four WM patients. B. WB analysis of GPER1 protein in a panel of six cancer cell lines (MCF7, MWCL-1, BCWM-1, DAUDI, RAJI, and MEC1). GAPDH was used as a loading control. C. CTG viability assay in BCWM-1 cells treated with indicated concentrations of GPER1 antagonists G15 and G-36. *Indicates p < 0.05 from a Student’s t-test. Ns indicates p > 0.05 from a Student’s t-test. Figure S3. A. WB analysis of p53, p21, BAX, and PUMA in primary CD19+ WM cells treated with G1 for 24 h. B. Wb analysis of Cyclin B1 in WM cell lines treated with indicated concentrations of G1. GAPDH was used as a loading control. C. Caspase 3/7 activity assay in WM cell lines treated with the indicated concentrations of G-1. Activity is represented relative to untreated cells. D. WB analysis of PARP, cleaved PARP, Caspase-3, and cleaved Caspase-3 in WM cell lines treated with the indicated concentrations of G-1. E. CTG viability assay and Caspase 3/7 activity assay in BCWM-1 cells, co-cultured for 48h with patient-derived bone marrow stromal cells and treated with G-1 [1 µM] or control. *Indicates p < 0.05 from a Student’s t-test. |
| doi_str_mv | 10.6084/m9.figshare.21087579 |
| format | Video |
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Analysis of GPER1 mRNA in public datasets GSE9656 (A) and GSE61597 (B). GSE9656: we analyzed GPER1 mRNA in CD19-selected peripheral blood B cells (pBCs; n = 7), bone marrow B cells from WM (WM-BCs; n = 12), bone marrow plasma cells from healthy donors (BM-PCs; n = 10), and WM plasma cells (WM-PCs, n = 9). GSE61597: we analyzed GPER1 mRNA in normal bone marrow CD25+ (n = 7) and CD25– (n = 9) B cells, clonal B cells from newly diagnosed patients with IgM MGUS (n=22), smoldering (n = 17), and symptomatic WM (n = 10). Figure S2. A. qRT-PCR analysis of GPER1 mRNA in MCF7 breast cancer cell line (positive control), MWCL-1 and BCWM-1 WM cell lines, and CD19+ primary cells from four WM patients. B. WB analysis of GPER1 protein in a panel of six cancer cell lines (MCF7, MWCL-1, BCWM-1, DAUDI, RAJI, and MEC1). GAPDH was used as a loading control. C. CTG viability assay in BCWM-1 cells treated with indicated concentrations of GPER1 antagonists G15 and G-36. *Indicates p < 0.05 from a Student’s t-test. Ns indicates p > 0.05 from a Student’s t-test. Figure S3. A. WB analysis of p53, p21, BAX, and PUMA in primary CD19+ WM cells treated with G1 for 24 h. B. Wb analysis of Cyclin B1 in WM cell lines treated with indicated concentrations of G1. GAPDH was used as a loading control. C. Caspase 3/7 activity assay in WM cell lines treated with the indicated concentrations of G-1. Activity is represented relative to untreated cells. D. WB analysis of PARP, cleaved PARP, Caspase-3, and cleaved Caspase-3 in WM cell lines treated with the indicated concentrations of G-1. E. CTG viability assay and Caspase 3/7 activity assay in BCWM-1 cells, co-cultured for 48h with patient-derived bone marrow stromal cells and treated with G-1 [1 µM] or control. *Indicates p < 0.05 from a Student’s t-test.</description><identifier>DOI: 10.6084/m9.figshare.21087579</identifier><language>eng</language><publisher>figshare</publisher><subject>Biochemistry ; Biological Sciences not elsewhere classified ; Cancer ; Cell Biology ; Chemical Sciences not elsewhere classified ; Developmental Biology ; FOS: Biological sciences ; FOS: Chemical sciences ; FOS: Clinical medicine ; Hematology ; Immunology ; Medicine ; Molecular Biology ; Pharmacology ; Physiology</subject><creationdate>2022</creationdate><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>776,1887</link.rule.ids><linktorsrc>$$Uhttps://commons.datacite.org/doi.org/10.6084/m9.figshare.21087579$$EView_record_in_DataCite.org$$FView_record_in_$$GDataCite.org$$Hfree_for_read</linktorsrc></links><search><creatorcontrib>Morelli, Eugenio</creatorcontrib><creatorcontrib>Hunter, Zachary R.</creatorcontrib><creatorcontrib>Fulciniti, Mariateresa</creatorcontrib><creatorcontrib>Gullà, Annamaria</creatorcontrib><creatorcontrib>Perrotta, Ida Daniela</creatorcontrib><creatorcontrib>Zuccalà, Valeria</creatorcontrib><creatorcontrib>Federico, Cinzia</creatorcontrib><creatorcontrib>Juli, Giada</creatorcontrib><creatorcontrib>Manzoni, Martina</creatorcontrib><creatorcontrib>Ronchetti, Domenica</creatorcontrib><creatorcontrib>Romeo, Enrica</creatorcontrib><creatorcontrib>Gallo Cantafio, Maria Eugenia</creatorcontrib><creatorcontrib>Soncini, Debora</creatorcontrib><creatorcontrib>Maltese, Lorenza</creatorcontrib><creatorcontrib>Rossi, Marco</creatorcontrib><creatorcontrib>Roccaro, Aldo M.</creatorcontrib><creatorcontrib>Cea, Michele</creatorcontrib><creatorcontrib>Tassone, Pierfrancesco</creatorcontrib><creatorcontrib>Neri, Antonino</creatorcontrib><creatorcontrib>Treon, Steven C.</creatorcontrib><creatorcontrib>Munshi, Nikhil C.</creatorcontrib><creatorcontrib>Viglietto, Giuseppe</creatorcontrib><creatorcontrib>Amodio, Nicola</creatorcontrib><title>Additional file 1 of Therapeutic activation of G protein-coupled estrogen receptor 1 in Waldenström Macroglobulinemia</title><description>Additional file 1: Figure S1. Analysis of GPER1 mRNA in public datasets GSE9656 (A) and GSE61597 (B). GSE9656: we analyzed GPER1 mRNA in CD19-selected peripheral blood B cells (pBCs; n = 7), bone marrow B cells from WM (WM-BCs; n = 12), bone marrow plasma cells from healthy donors (BM-PCs; n = 10), and WM plasma cells (WM-PCs, n = 9). GSE61597: we analyzed GPER1 mRNA in normal bone marrow CD25+ (n = 7) and CD25– (n = 9) B cells, clonal B cells from newly diagnosed patients with IgM MGUS (n=22), smoldering (n = 17), and symptomatic WM (n = 10). Figure S2. A. qRT-PCR analysis of GPER1 mRNA in MCF7 breast cancer cell line (positive control), MWCL-1 and BCWM-1 WM cell lines, and CD19+ primary cells from four WM patients. B. WB analysis of GPER1 protein in a panel of six cancer cell lines (MCF7, MWCL-1, BCWM-1, DAUDI, RAJI, and MEC1). GAPDH was used as a loading control. C. CTG viability assay in BCWM-1 cells treated with indicated concentrations of GPER1 antagonists G15 and G-36. *Indicates p < 0.05 from a Student’s t-test. Ns indicates p > 0.05 from a Student’s t-test. Figure S3. A. WB analysis of p53, p21, BAX, and PUMA in primary CD19+ WM cells treated with G1 for 24 h. B. Wb analysis of Cyclin B1 in WM cell lines treated with indicated concentrations of G1. GAPDH was used as a loading control. C. Caspase 3/7 activity assay in WM cell lines treated with the indicated concentrations of G-1. Activity is represented relative to untreated cells. D. WB analysis of PARP, cleaved PARP, Caspase-3, and cleaved Caspase-3 in WM cell lines treated with the indicated concentrations of G-1. E. 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Analysis of GPER1 mRNA in public datasets GSE9656 (A) and GSE61597 (B). GSE9656: we analyzed GPER1 mRNA in CD19-selected peripheral blood B cells (pBCs; n = 7), bone marrow B cells from WM (WM-BCs; n = 12), bone marrow plasma cells from healthy donors (BM-PCs; n = 10), and WM plasma cells (WM-PCs, n = 9). GSE61597: we analyzed GPER1 mRNA in normal bone marrow CD25+ (n = 7) and CD25– (n = 9) B cells, clonal B cells from newly diagnosed patients with IgM MGUS (n=22), smoldering (n = 17), and symptomatic WM (n = 10). Figure S2. A. qRT-PCR analysis of GPER1 mRNA in MCF7 breast cancer cell line (positive control), MWCL-1 and BCWM-1 WM cell lines, and CD19+ primary cells from four WM patients. B. WB analysis of GPER1 protein in a panel of six cancer cell lines (MCF7, MWCL-1, BCWM-1, DAUDI, RAJI, and MEC1). GAPDH was used as a loading control. C. CTG viability assay in BCWM-1 cells treated with indicated concentrations of GPER1 antagonists G15 and G-36. *Indicates p < 0.05 from a Student’s t-test. Ns indicates p > 0.05 from a Student’s t-test. Figure S3. A. WB analysis of p53, p21, BAX, and PUMA in primary CD19+ WM cells treated with G1 for 24 h. B. Wb analysis of Cyclin B1 in WM cell lines treated with indicated concentrations of G1. GAPDH was used as a loading control. C. Caspase 3/7 activity assay in WM cell lines treated with the indicated concentrations of G-1. Activity is represented relative to untreated cells. D. WB analysis of PARP, cleaved PARP, Caspase-3, and cleaved Caspase-3 in WM cell lines treated with the indicated concentrations of G-1. E. CTG viability assay and Caspase 3/7 activity assay in BCWM-1 cells, co-cultured for 48h with patient-derived bone marrow stromal cells and treated with G-1 [1 µM] or control. *Indicates p < 0.05 from a Student’s t-test.</abstract><pub>figshare</pub><doi>10.6084/m9.figshare.21087579</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Biochemistry Biological Sciences not elsewhere classified Cancer Cell Biology Chemical Sciences not elsewhere classified Developmental Biology FOS: Biological sciences FOS: Chemical sciences FOS: Clinical medicine Hematology Immunology Medicine Molecular Biology Pharmacology Physiology |
| title | Additional file 1 of Therapeutic activation of G protein-coupled estrogen receptor 1 in Waldenström Macroglobulinemia |
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