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. Author manuscript; available in PMC: 2020 May 27.
Published in final edited form as: Nat Chem Biol. 2019 Jan 14;15(2):179–188. doi: 10.1038/s41589-018-0200-7

Fig. 3 |. Generation and characterization of a NVS-ZP7–4 compound-resistant cell line.

Fig. 3 |

a, Comparison of effects of NVS-ZP7–4 on apoptosis/cell death in the parental TALL-1 and resistant TALL-1 cell line (TLR1). Dose response of NVS-ZP7–4 following 72 h of compound treatment. Data shown are the sum of percentage dead and apoptotic cells by annexin V/propidium iodide staining after compound treatment. Each bar graph represents the data from one individual experiment in which two independent samples are treated with compound. The average readout value for these samples is represented by the dot-plot bar graph. Each experiment was performed three independent times. b, Levels of Notch3 intracellular domain (ICD3) in the parental (TALL-1) and compound-resistant TALL-1 (TLR1) cell line following 20 h of 1 μM NVS-ZP7–4 or 10 μM DAPT treatment. Full length gels are shown in Supplementary Fig. 13, and the experiment was performed two independent times with representative western blot data shown. c, Schematic of sequencing and mutation detection in compound-resistant cell line. NGS, next generation sequencing. d, Effect of siRNA knockdown of genes identified from resistant cell line sequencing in combination with 20 nM NVS-ZP7–4 in ERSE reporter gene assay. Plate median normalized ERSE-Luc data are shown on the y axis.