Gapmer-mediated silencing of mutTNPO3 as a therapeutic strategy for LGMDD2

This dataset contains experimental data generated to evaluate the silencing of mutant TNPO3 (TNPO3mut) using antisense gapmer oligonucleotides in an immortalized cell line derived from patients with Limb-Girdle Muscular Dystrophy D2 (LGMDD2). A healthy control immortalized cell line (AB1190) was used as control. Experiments were performed under the following conditions: untreated control cells, untreated LGMDD2 cells, LGMDD2 cells treated with gapmer 1 or gapmer 2 at 100 nM or 200 nM, and cells treated with a control gapmer, referred to in the files as cGP or SC. The dataset includes the following types of data: Jess (capillary-based immunoassay) files assessing the expression of TNPO3mut and total TNPO3 in LGMDD2 cells treated with gapmers. Jess files evaluating TNPO3 expression in AB1190 control cells treated with gapmers to assess allele specificity toward the mutant transcript. Cell proliferation assays performed in LGMDD2 cells treated with gapmers to evaluate potential cytotoxicity. Immunofluorescence (IF) images of desmin staining to assess whether gapmer treatment rescues differentiation defects in LGMDD2 cells. Immunofluorescence images of mutant TNPO3 after treatment with Cy3-labeled gapmer 2. Expression analysis of MBNL1 following gapmer treatment. Immunofluorescence images of MBNL1 at T0 to evaluate its baseline subcellular distribution. Immunofluorescence images of MBNL1 at T7 to assess changes in subcellular localization and potential rescue after gapmer treatment. Confocal microscopy images of LysoTracker staining to study autophagy-related alterations following gapmer treatment. Western blot analyses of LC3 and p62 to further evaluate autophagy-related pathways after gapmer treatment. RNA-seq–derived counts per million (CPM) of TNPO3 alleles (WT and mutant) across experimental conditions. Agarose gel images of RT-PCR analysis of MBNL1-dependent alternative splicing events.