Introduction
- Vancheri C.
- Kreuter M.
- Richeldi L.
- Ryerson C.J.
- Valeyre D.
- Grutters J.C.
- Wiebe S.
- Stansen W.
- Quaresma M.
- Stowasser S.
- Wuyts W.A.
Nintedanib with add-on pirfenidone in idiopathic pulmonary fibrosis: results of the in journey trial.
- Hellemans J.
- Preobrazhenska O.
- Willaert A.
- Debeer P.
- Verdonk P.C.
- Costa T.
- Janssens K.
- Menten B.
- Van Roy N.
- Vermeulen S.J.
- Savarirayan R.
- Van Hul W.
- Vanhoenacker F.
- Huylebroeck D.
- De Paepe A.
- et al.
- Hellemans J.
- Preobrazhenska O.
- Willaert A.
- Debeer P.
- Verdonk P.C.
- Costa T.
- Janssens K.
- Menten B.
- Van Roy N.
- Vermeulen S.J.
- Savarirayan R.
- Van Hul W.
- Vanhoenacker F.
- Huylebroeck D.
- De Paepe A.
- et al.
- Hellemans J.
- Preobrazhenska O.
- Willaert A.
- Debeer P.
- Verdonk P.C.
- Costa T.
- Janssens K.
- Menten B.
- Van Roy N.
- Vermeulen S.J.
- Savarirayan R.
- Van Hul W.
- Vanhoenacker F.
- Huylebroeck D.
- De Paepe A.
- et al.
Results
Fibroblast TGFβ responsiveness is potentiated by ECM stiffness and inhibited by LEMD3 expression

- Hellemans J.
- Preobrazhenska O.
- Willaert A.
- Debeer P.
- Verdonk P.C.
- Costa T.
- Janssens K.
- Menten B.
- Van Roy N.
- Vermeulen S.J.
- Savarirayan R.
- Van Hul W.
- Vanhoenacker F.
- Huylebroeck D.
- De Paepe A.
- et al.
LEMD3–SMAD2/3 complexes are inversely correlated to substrate stiffness and occur throughout the cell

LEMD3–Smad2/3 complexes are inhibited by actin polymerization and not potentiated by disrupting cytoplasmic–nuclear–lamina–LEMD3 coupling

LEMD3 fragments are generated by a serine protease and differentially regulated by the integrity of the nuclear lamina

C-terminal fragment of LEMD3 binds Smad2/3 in a stiffness-dependent fashion, antagonizes Smad2/3–Smad4 complexes, and binds PPM1α

LEMD3–SMAD2/3 complexes are more cytosolic, and their frequencies are more varied in IPF biopsies than non-IPF biopsies

Discussion
- Vancheri C.
- Kreuter M.
- Richeldi L.
- Ryerson C.J.
- Valeyre D.
- Grutters J.C.
- Wiebe S.
- Stansen W.
- Quaresma M.
- Stowasser S.
- Wuyts W.A.
Nintedanib with add-on pirfenidone in idiopathic pulmonary fibrosis: results of the in journey trial.
- DePianto D.J.
- Chandriani S.
- Abbas A.R.
- Jia G.
- N′Diaye E.N.
- Caplazi P.
- Kauder S.E.
- Biswas S.
- Karnik S.K.
- Ha C.
- Modrusan Z.
- Matthay M.A.
- Kukreja J.
- Collard H.R.
- Egen J.G.
- et al.

- Hellemans J.
- Preobrazhenska O.
- Willaert A.
- Debeer P.
- Verdonk P.C.
- Costa T.
- Janssens K.
- Menten B.
- Van Roy N.
- Vermeulen S.J.
- Savarirayan R.
- Van Hul W.
- Vanhoenacker F.
- Huylebroeck D.
- De Paepe A.
- et al.
Experimental procedures
Primary fibroblast cultures and transfection, human lung biopsies, and other reagents
Generation of recombinant constructs
DN LEM construct
Western blotting
qPCR
Smad-driven luciferase assays
Proximity ligation assays
Ex vivo PLA
In vitro PLA
Atomic force microscopy and cell morphology
Mass spectroscopy on LEMD3 fragments
Author contributions
Acknowledgments
Supplementary Material
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Footnotes
This work was supported, in whole or in part, by National Institutes of Health Grants R01 HL127283 and R01 HL132585 (to T. H. B.) and F30 HL122065 (to Dwight M. C.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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