@article{4bd0405215324001b574eadec18f999c,
title = "Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function",
abstract = "Upon initial immune challenge, dendritic cells (DCs) migrate to lymph nodes and interact with fibroblastic reticular cells (FRCs) via C-type lectin-like receptor 2 (CLEC-2). CLEC-2 binds to the membrane glycoprotein podoplanin (PDPN) on FRCs, inhibiting actomyosin contractility through the FRC network and permitting lymph node expansion. The hyaluronic acid receptor CD44 is known to be required for FRCs to respond to DCs but the mechanism of action is not fully elucidated. Here, we use DNA-PAINT, a quantitative single molecule super-resolution technique, to visualize and quantify how PDPN clustering is regulated in the plasma membrane of FRCs. Our results indicate that CLEC-2 interaction leads to the formation of large PDPN clusters (i.e. more than 12 proteins per cluster) in a CD44-dependent manner. These results suggest that CD44 expression is required to stabilize large pools of PDPN at the membrane of FRCs upon CLEC-2 interaction, revealing the molecular mechanism through which CD44 facilitates cellular crosstalk between FRCs and DCs.",
keywords = "CD44, CLEC-2, DNA-PAINT, podoplanin, qPAINT, super-resolution",
author = "Lim, {Shu En} and Joseph, {Megan D.} and {de Winde}, {Charlotte M.} and Acton, {Sophie E.} and Sabrina Simoncelli",
note = "Funding Information: S.E.L. is supported by the Wellcome Trust (grant no. 225439/Z/22/Z). M.D.J. is supported by the BBSRC (grant no. BB/T008709/1). C.M.d.W. is supported by the Dutch Research Foundation (Nederlandse Organisatie voor Wetenschappelijk Onderzoek, NWO) via a Rubicon Postdoctoral Fellowship (019.162LW.004 to C.M.d.W.), European Research Council Starting Grant (grant no. LNEXPANDS to S.E.A.). S.E.A. acknowledges financial support from a European Research Council Starting Grant (grant no. LNEX-PANDS), a Cancer Research UK Careers Development Fellowship (grant no. CRUK-A19763) and the Medical Research Council (grant no. MC-U12266B). S.S. acknowledges financial support from the Royal Society through a Dorothy Hodgkin (grant no. DHF\R1\191019) fellowship and a Research Fellows Enhanced Research Expenses (grant no. RF\ERE\210227). Publisher Copyright: {\textcopyright} 2023 The Authors.",
year = "2023",
month = may,
day = "10",
doi = "https://doi.org/10.1098/rsob.220377",
language = "English",
volume = "13",
journal = "Open biology",
issn = "2046-2441",
publisher = "The Royal Society",
number = "5",
}