Targeting DMPK with Antisense Oligonucleotide Improves Muscle Strength in Myotonic Dystrophy Type 1 Mice Dominic Jauvin, Jessina Chrétien, Sanjay K. Pandey, Laurie Martineau, Lucille Revillod, Guillaume Bassez, Aline Lachon, A. Robert McLeod, Geneviève Gourdon, Thurman M. Wheeler, Charles A. Thornton, C. Frank Bennett, and Jack Puymirat
Supplementary Material
Figure S1 – Foci reduction by ASOs FISH on DM1 myotubes transfected with ASOs. a. DM1 shows a great abundance of nuclear retained DMPK mRNA. b.c. Transfection with ASOs greatly reduces the presence of nuclear foci. Scale bar = 20 µm.
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Figure S2 – Redistribution of MBNL1 after treatment with ASOs. MBNL1 immunostaining combined with FISH in myotubes. a. Normal myotubes do not contain DMPK induced foci and MBNL1 is homogeneously distributed inside nuclei. b. DM1 myotubes shows sequestration of MBNL1 with nuclear foci and loss of soluble MBNL1. c.d. ASOs transfected in DM1 myotubes induces degradation of expanded DMPK mRNA, which enables MBNL1 redistribution throughout the nuclei. Scale bar = 20 µm.
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Figure S3 – Necrosis and apoptosis assays on normal cells treated by ASOs. No significant effect of ASOs on necrosis (a) and apoptosis (b) after 48 hours of treatment (n =2).
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Figure S4 - Foci reduction in DMSXL mice quadriceps. FISH on transverse section of DMSXL mice quadriceps. Systemic injection of ASOs produces high enough intracellular levels in muscle to reduce the presence of nuclear foci. ISIS 486178 (cEt chemistry) shows a stronger effect (b) than ISIS 445569 (MOE chemistry) (c) at half the dose (DMSXL n=3; ASOs n=9). Scale bar = 5 µm.
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Figure S5 – Systemic administration of ISIS 486178 in wild-type mice. ISIS 486178 targets not only the DMPK mRNA but also the Dmpk mRNA as shown by systemic injection (25 mg/kg) in wild-type mice (saline WT n = 7, treated WT n = 8). * p
