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(A) Schematic illustrations of the establishment of CNDAC-resistant HL-60 and PL-21 cells by step-wise increasing drug concentrations during cell culture and of the establishment of single cell-derived clones by limited dilution. Moreover, representative Western blots indicating SAMHD1 and DCK levels in CNDAC-adapted HL-60 (HL-60 r CNDACI-XII) and PL21 (PL21 r CNDACI-XII) sublines and in single cell-derived clonal sublines of these cell lines. GAPDH and β-Actin served as loading controls. (B) Resistance profiles of CNDAC-adapted HL-60 and PL-21 sublines and single cell-derived clones of HL-60 and PL-21. Upper spider webs show sensitivity to the cytotoxic drugs CNDAC, 6-Thioguanine (6-TG), Clofarabine (CLOF), Cladribine (CLAD), Fludarabine (FLU), Gemcitabine (GEM), Decitabine (DAC), 5-Azacytidine (AZA), Daunorubicin (DAU), Cytarabine (ARA-C), and <t>Sapacitabine</t> (SAP), while lower spider webs display sensitivity to the targeted drugs Vismodegib (VISMO), Olaparib (OLA), Ganetespib (GANE), Gedatolisib (GEDA), Volasertib (VOLA), Molibresib (MOLI), and Venetoclax (VENE). Values are depicted as fold changes in drug concentrations that reduce cell viability by 50% (IC 50 s) between the respective parental AML cell line (shown in red) and the resistant cell lines or clones. Points closer to the centre than red lines indicate higher sensitivity to drugs in CNDAC-resistant sublines or clonal sublines than in parental cell lines, while points lying outside red lines indicate reduced sensitivity to the respective drug. IC 50 fold changes are shown as means from three independent experiments. Numerical values are provided in Supplementary Table 6.
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(A) Schematic illustrations of the establishment of CNDAC-resistant HL-60 and PL-21 cells by step-wise increasing drug concentrations during cell culture and of the establishment of single cell-derived clones by limited dilution. Moreover, representative Western blots indicating SAMHD1 and DCK levels in CNDAC-adapted HL-60 (HL-60 r CNDACI-XII) and PL21 (PL21 r CNDACI-XII) sublines and in single cell-derived clonal sublines of these cell lines. GAPDH and β-Actin served as loading controls. (B) Resistance profiles of CNDAC-adapted HL-60 and PL-21 sublines and single cell-derived clones of HL-60 and PL-21. Upper spider webs show sensitivity to the cytotoxic drugs CNDAC, 6-Thioguanine (6-TG), Clofarabine (CLOF), Cladribine (CLAD), Fludarabine (FLU), Gemcitabine (GEM), Decitabine (DAC), 5-Azacytidine (AZA), Daunorubicin (DAU), Cytarabine (ARA-C), and Sapacitabine (SAP), while lower spider webs display sensitivity to the targeted drugs Vismodegib (VISMO), Olaparib (OLA), Ganetespib (GANE), Gedatolisib (GEDA), Volasertib (VOLA), Molibresib (MOLI), and Venetoclax (VENE). Values are depicted as fold changes in drug concentrations that reduce cell viability by 50% (IC 50 s) between the respective parental AML cell line (shown in red) and the resistant cell lines or clones. Points closer to the centre than red lines indicate higher sensitivity to drugs in CNDAC-resistant sublines or clonal sublines than in parental cell lines, while points lying outside red lines indicate reduced sensitivity to the respective drug. IC 50 fold changes are shown as means from three independent experiments. Numerical values are provided in Supplementary Table 6.

Journal: bioRxiv

Article Title: Differences between intrinsic and acquired nucleoside analogue resistance in acute myeloid leukaemia cells

doi: 10.1101/2021.07.19.452885

Figure Lengend Snippet: (A) Schematic illustrations of the establishment of CNDAC-resistant HL-60 and PL-21 cells by step-wise increasing drug concentrations during cell culture and of the establishment of single cell-derived clones by limited dilution. Moreover, representative Western blots indicating SAMHD1 and DCK levels in CNDAC-adapted HL-60 (HL-60 r CNDACI-XII) and PL21 (PL21 r CNDACI-XII) sublines and in single cell-derived clonal sublines of these cell lines. GAPDH and β-Actin served as loading controls. (B) Resistance profiles of CNDAC-adapted HL-60 and PL-21 sublines and single cell-derived clones of HL-60 and PL-21. Upper spider webs show sensitivity to the cytotoxic drugs CNDAC, 6-Thioguanine (6-TG), Clofarabine (CLOF), Cladribine (CLAD), Fludarabine (FLU), Gemcitabine (GEM), Decitabine (DAC), 5-Azacytidine (AZA), Daunorubicin (DAU), Cytarabine (ARA-C), and Sapacitabine (SAP), while lower spider webs display sensitivity to the targeted drugs Vismodegib (VISMO), Olaparib (OLA), Ganetespib (GANE), Gedatolisib (GEDA), Volasertib (VOLA), Molibresib (MOLI), and Venetoclax (VENE). Values are depicted as fold changes in drug concentrations that reduce cell viability by 50% (IC 50 s) between the respective parental AML cell line (shown in red) and the resistant cell lines or clones. Points closer to the centre than red lines indicate higher sensitivity to drugs in CNDAC-resistant sublines or clonal sublines than in parental cell lines, while points lying outside red lines indicate reduced sensitivity to the respective drug. IC 50 fold changes are shown as means from three independent experiments. Numerical values are provided in Supplementary Table 6.

Article Snippet: CNDAC was purchased from biorbyt (via Biozol, Eching, Germany), 5-azacytidine, cytarabine, cladribine, clofarabine, decitabine, and fludarabine from Tocris Biosciences (via Bio-Techne GmbH, Wiesbaden, Germany), 6-thioguanine, ganetespib, molibresib, olaparib, sapacitabine, venetoclax, and vismodegib from MedChemExpress (via Hycultec, Beutelsbach, Germany), daunorubicin, gedatolisib, and volasertib from Selleckchem (Berlin, Germany), gemcitabine from Hexal (Holzkirchen, Germany), GTP and dATP from Thermo Scientific (Dreieich, Germany), and CNDAC-TP from Jena Bioscience GmbH (Jena, Germany).

Techniques: Cell Culture, Derivative Assay, Clone Assay, Western Blot