Supplementary Materials

Supplementary Material for:

ACVR1R206H receptor mutation causes fibrodysplasia ossificans progressiva by imparting responsiveness to activin A

Sarah J. Hatsell, Vincent Idone, Dana M. Alessi Wolken, Lily Huang, Hyon J. Kim, Lili Wang, Xialing Wen, Kalyan C. Nannuru, Johanna Jimenez, Liqin Xie, Nanditha Das, Genevieve Makhoul, Rostislav Chernomorsky, David D'Ambrosio, Richard A. Corpina, Christopher J. Schoenherr, Kieran Feeley, Paul B. Yu, George D. Yancopoulos, Andrew J. Murphy, Aris N. Economides*

*Corresponding author. E-mail: aris{at}regeneron.com

Published 2 September 2015, Sci. Transl. Med. 7, 303ra137 (2015)
DOI: 10.1126/scitranslmed.aac4358

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Expression of ACVR1 in HEK293/BRE-Luc cells is detected by surface staining.
  • Fig. S2. BMP family ligands parse into two classes with respect to the response they elicit in ACVR1-overexpressing compared to ACVR1[R206H]-overexpressing lines.
  • Fig. S3. Acvr1 exon 5 can be skipped during splicing of the Acvr1[R206H]FlEx pre-mRNA.
  • Fig. S4. Activin A–induced Smad2/3 signaling in ACVR1[R206H]-expressing cells is similar to wild-type ES cells.
  • Fig. S5. FKBP12 inhibition by FK506 does not render wild-type ACVR1 into an activin-responsive receptor.
  • Fig. S6. Activins and BMPs bind to wild-type ACVR1 and form complexes with ACVR2A.
  • Fig. S7. Acvr1[R206H]FlEx/+; Gt(ROSA26)SorCreERT2/+ mice develop HO as early as 2 weeks after tamoxifen administration.
  • Fig. S8. Quantification of HO formation Acvr1[R206H]FlEx/+; Gt(ROSA26)SorCreERT2/+ mice from 2 to 6 weeks after tamoxifen administration.
  • Fig. S9. H&E sections from HO lesions shows a variety of different cell types, mirroring those found in FOP lesions.
  • Fig. S10. ACVR2A-Fc and ACVR2B-Fc prevent ectopic bone formation in Acvr1[R206H]FlEx/+; Gt(ROSA26)SorCreERT2/+ mice.
  • Fig. S11. Activin A causes HO in tamoxifen-treated Acvr1[R206H]FlEx/+; Gt(ROSA26)SorCreERT2/+ but not wild-type mice.
  • Fig. S12. Tamoxifen does not alter serum activin A levels.
  • Fig. S13. Anti–activin A blocking mAb blocks the activity of inhibin A–containing activins but not activin B.
  • Fig. S14. In mouse ES cells, the Acvr1[R206H]FlEx allele is inverted by 24 hours after 100 nM tamoxifen treatment.
  • Table S1. Number of mice with HO lesions in treatment experiments.
  • Table S2. Acvr1[R206H]FlEx/+ mice cannot be bred to homozygosity.

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