Figure S3. Ex7 skipping-antisense oligonucleotides sequence and characterization.

(A) Genomic sequence of the highly conserved 36-bp exons of MBNL1, MBNL2, and MBNL3 (in orange) with the surrounding intronic region. Above is shown the pairing position of the tree AONs directed against MBNL1 ex7. Below, the conservation index of the nucleotidic sequence. (B) PSI of MBNL1 ex7 (blue), MBNL2 ex7 (green), and MBNL3 exon 6 upon 100 nM AON transfection at 48 h in PC3 cells. (C) FLA-PCR visualization of MBNL1 cDNA isoforms upon AON #592 transfection in LnCap cells (100 nM, at 48 h posttransfection). On the X axis is represented the nucleotide size and on the Y axis, the fluorescence intensity of the amplicon. Data are representative of multiple independent experiments. (D) Immunofluorescence of PC3 cells upon transfection with 100 nM of AON SCR and MBNL1 at 48 h posttransfection. (E) PSI of MBNL1 ex7 in different cell lines upon transfection of AON MBNL1 at different concentrations. Cells were collected at 48 h. (F) AON SCR–normalized PC3 cell viability at different AON MBNL1 concentrations. Cells were counted at 48 h. (G) PSI of MBNL1 ex7 (black) and ex5 (gray) in PC3 and LnCap cells after 100 nM AON transfection. Cells were collected at 48 h. (H) Cell number of PC3 and LnCap cells after 100 nM AON transfection. Cells were counted at 24 h. (I) Quantification of western blot band intensity of MBNL1 from Fig 3C. (J) Ex7 PSI (blue bars), overall MBNL1 relative mRNA expression (green bars), and relative cell count of LnCap cells transfected with AON SCR, AON MBNL1 (100 nM), siRNA Scrambled, and siRNA MBNL1 (25 nM). (K) FLA-PCR profile of AON and siRNA transfection in PC3 cells.LSA-2018-00157_FigS3.pdf ^{(9.3MB, pdf)}