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A mutant-cell library for systematic analysis of heparan sulfate structure-function relationships.

Nature methods | 2018

Heparan sulfate (HS) is a complex linear polysaccharide that modulates a wide range of biological functions. Elucidating the structure-function relationship of HS has been challenging. Here we report the generation of an HS-mutant mouse lung endothelial cell library by systematic deletion of HS genes expressed in the cell. We used this library to (1) determine that the strictly defined fine structure of HS, not its overall degree of sulfation, is more important for FGF2-FGFR1 signaling; (2) define the epitope features of commonly used anti-HS phage display antibodies; and (3) delineate the fine inter-regulation networks by which HS genes modify HS and chain length in mammalian cells at a cell-type-specific level. Our mutant-cell library will allow robust and systematic interrogation of the roles and related structures of HS in a cellular context.

Pubmed ID: 30377379 RIS Download

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Associated grants

  • Agency: NHLBI NIH HHS, United States
    Id: P01 HL107150
  • Agency: NHLBI NIH HHS, United States
    Id: R21 HL131553
  • Agency: NHLBI NIH HHS, United States
    Id: R01 HL093339
  • Agency: NIGMS NIH HHS, United States
    Id: P41 GM103390
  • Agency: NCI NIH HHS, United States
    Id: U01 CA225784
  • Agency: NIA NIH HHS, United States
    Id: R56 AG062344
  • Agency: NHLBI NIH HHS, United States
    Id: P01 HL131474

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FlowJo (tool)

RRID:SCR_008520

Software for single-cell flow cytometry analysis. Its functions include management, display, manipulation, analysis and publication of the data stream produced by flow and mass cytometers.

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C57BL/6J (tool)

RRID:IMSR_JAX:000664

Mus musculus with name C57BL/6J from IMSR.

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