Quantitative analysis of PARP inhibitor toxicity by multidimensional fluorescence microscopy
Current methods employed to evaluate PARP inhibitor (PARPi) toxicity can be time consuming, have limited sensitivity, may not be well suited for screening purposes, or yield results from cell population averages rather than single cell information. To overcome these limitations we provide a detailed protocol to assess PARPi toxicity at multiple phenotypic levels based on a high-content microscopy workflow. This approach takes two days and allows for sensitive cell cycle resolved analyses of PARPi-evoked DNA damage response signaling and PARP trapping. The approach can be applied to evaluate conditions of PARPi hypersensitivity and resistance, and to obtain insights in the cellular mechanisms of drug synergism. Moreover, the described workflow is compatible with high-throughput screening for the discovery and characterization of functional interactions between PARPi and other clinically relevant drugs.
Figure 1
Posted 28 Jan, 2019
Quantitative analysis of PARP inhibitor toxicity by multidimensional fluorescence microscopy
Posted 28 Jan, 2019
Current methods employed to evaluate PARP inhibitor (PARPi) toxicity can be time consuming, have limited sensitivity, may not be well suited for screening purposes, or yield results from cell population averages rather than single cell information. To overcome these limitations we provide a detailed protocol to assess PARPi toxicity at multiple phenotypic levels based on a high-content microscopy workflow. This approach takes two days and allows for sensitive cell cycle resolved analyses of PARPi-evoked DNA damage response signaling and PARP trapping. The approach can be applied to evaluate conditions of PARPi hypersensitivity and resistance, and to obtain insights in the cellular mechanisms of drug synergism. Moreover, the described workflow is compatible with high-throughput screening for the discovery and characterization of functional interactions between PARPi and other clinically relevant drugs.
Figure 1
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