期刊
METHODS
卷 193, 期 -, 页码 38-45出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2020.05.004
关键词
Receptor tyrosine kinases; Fibroblast growth factor receptor; FGFR network; Single-molecule localization microscopy; DNA-PAINT; Exchange DNA-PAINT
资金
- State of Hesse (LOEWE)
- State of Hesse (DynaMem)
- State of Hesse (FCI)
- German Science Foundation [SFB 1177]
The study established an imaging and analysis pipeline for multiplexed single-molecule localization microscopy (SMLM) of the FGFR network at the plasma membrane. From the super-resolution imaging data, information on FGFR density, spatial distribution, and inner-subfamily colocalization was extracted.
Fibroblast growth factor receptors (FGFRs) are a subfamily of receptor tyrosine kinases and central players in health and disease. Following ligand binding and the formation of homo- and heteromeric complexes, FGFRs initiate a cellular response. Challenges in studying FGFR activation are inner-subfamily interactions and a complex heterogeneity of these in the cell membrane, which demand for observation techniques that can resolve individual protein complexes and that are compatible with endogenous protein levels. Here, we established an imaging and analysis pipeline for multiplexed single-molecule localization microscopy (SMLM) of the FGFR network at the plasma membrane. Using DNA-labeled primary antibodies, we visualize all four FGFRs in the same cell with near-molecular spatial resolution. From the super-resolution imaging data, we extract information on FGFR density, spatial distribution, and inner-subfamily colocalization. Our approach is straightforward and easily adaptable to other multiplexed SMLM data of membrane proteins.
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