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PALP: A rapid imaging technique for stratifying ferroptosis sensitivity in normal and tumor tissues in situ

Cell Chem Biol. 2021-11; 
Fengxiang Wang, Emily T Graham, Nathchar Naowarojna, Zhennan Shi, Yuqi Wang, Guanglei Xie, Lili Zhou, Wendy Salmon, Jie-Min Jia, Xi Wang, Yuwei Huang, Stuart L Schreiber, Yilong Zou
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Proteins, Expression, Isolation and Analysis … Calibrated samples were diluted with 4x lithium dodecyl sulfate (LDS) sample buffer (Novus), separated by SDS–PAGE using SurePAGE 4–20% Bis-Tris protein gels (GenScript), and transferred to 0.45 μm PVDF membranes by an eBlot L1 protein-transfer system (Genscript). … Get A Quote

摘要

Ferroptosis is an emerging cancer suppression strategy. However, how to select cancer patients for treating with ferroptosis inducers remains challenging. Here, we develop photochemical activation of membrane lipid peroxidation (PALP), which uses targeted lasers to induce localized polyunsaturated fatty acyl (PUFA)-lipid peroxidation for reporting ferroptosis sensitivity in cells and tissues. PALP captured by BODIPY-C11 can be suppressed by lipophilic antioxidants and iron chelation, and is dependent on PUFA-lipid levels. Moreover, we develop PALPv2, for studying lipid peroxidation on selected membranes along the z axis in live cells using two-photon microscopes. Using PALPv1, we detect PUFA-lipids in multiple ... More

关键词

cancer, ferroptosis sensitivity stratification, imaging, in situ quantitation, lipid peroxidation, polyunsaturated lipid