Supplementary Materials

Supplementary Material for:

Adenylyl cyclase activating polypeptide reduces phosphorylation and toxicity of the polyglutamine-expanded androgen receptor in spinobulbar muscular atrophy

Maria Josè Polanco, Sara Parodi, Diana Piol, Conor Stack, Mathilde Chivet, Andrea Contestabile, Helen C. Miranda, Patricia M.-J. Lievens, Stefano Espinoza, Tobias Jochum, Anna Rocchi, Christopher Grunseich, Raul R. Gainetdinov, Andrew C. B. Cato, Andrew P. Lieberman, Albert R. La Spada, Fabio Sambataro, Kenneth H. Fischbeck, Illana Gozes, Maria Pennuto*

*Corresponding author. Email: mpennuto{at}dti.telethon.it

Published 21 December 2016, Sci. Transl. Med. 8, 370ra181 (2016)
DOI: 10.1126/scitranslmed.aaf9526

This PDF file includes:

  • Table S1. List of PACAP analogs and cAMP release in MN-1 cells treated with either PACAP or the indicated PACAP analogs.
  • Fig. S1. Overexpression of PKA reduces the accumulation of the upper isoform of polyQ-AR.
  • Fig. S2. PACAP stimulates the release of cAMP in MN-1 cells expressing AR24Q and AR65Q.
  • Fig. S3. VIP does not modify the accumulation of the upper isoform of polyQ-AR.
  • Fig. S4. Expression of polyQ-AR in MN-1 cells results in caspase 3 activation.
  • Fig. S5. Analysis of polyQ-AR phosphorylation upon inhibition of specific cellular phosphatases and kinases.
  • Fig. S6. Modulation of CDK1 and CDK5 activity does not affect the accumulation of the upper isoform of polyQ-AR.
  • Fig. S7. Nonexpanded AR and polyQ-AR colocalize with endogenous CDK2 in the absence and presence of androgens in MN-1 and PC12 cells.
  • Fig. S8. Nonexpanded AR and polyQ-AR colocalize with endogenous CDK2 in the brainstem motor neurons of control and knock-in SBMA mice.
  • Fig. S9. Nonexpanded AR and polyQ-AR colocalize with endogenous CDK2 in the motor neurons of the lumbar spinal cord of control and knock-in SBMA mice.
  • Fig. S10. Nonexpanded AR colocalizes with endogenous CDK2 in the quadriceps of control mice.
  • Fig. S11. Endogenous nonexpanded AR and polyQ-AR colocalize with endogenous CDK2 in control and SBMA patient-derived motor neurons and NPCs.
  • Fig. S12. Endogenous polyQ-AR colocalizes with endogenous CDK2 in the spinal cord of an SBMA patient.
  • Fig. S13. Forskolin, PACAP, and the pan-phosphatase inhibitor OA stimulate p21Cip1 expression.
  • Fig. S14. Ser96 of AR is conserved throughout evolution.
  • Fig. S15. Analysis of phosphoresistant polyQ-AR variants.
  • Fig. S16. Ser96 phosphorylation is responsible for the formation of the upper isoform of polyQ-AR.
  • Fig. S17. Phosphodefective and phosphomimetic substitution of Ser96 does not affect polyQ-AR subcellular localization.
  • Fig. S18. Forskolin and PACAP, and not VIP, reduce polyQ-AR aggregation.
  • Fig. S19. Activation of the AC/PKA signaling increases the turnover of polyQ-AR.
  • Fig. S20. OA reduces the accumulation of phosphorylated polyQ-AR by inducing degradation through the UPS.
  • Fig. S21. PACAP induces cAMP production in vivo.
  • Fig. S22. Effect of intranasal administration of peptide 7 on body weight, food intake, and survival of AR113Q mice.
  • Fig. S23. Peptide 7 increases the CSA of glycolytic fibers in the skeletal muscle of AR113Q mice and restores the expression of glycolytic genes to normal levels.
  • Fig. S24. Intranasal administration of peptide 7 decreases polyQ-AR phosphorylation and accumulation in SBMA mice without altering CDK2, p27Kip1, and p57Kip2 expression levels.
  • Fig. S25. Data presented for experiments with sample sizes of less than 20.

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