Fig. 5: A1155463 shows senolytic activity on DM1 primary myoblasts culture.

a Cell viability assay of A1155463 at different concentrations using control (Ctrl) and DM1 patients’ myoblasts (n = 5). *p = 0.038 (100 nM), *p = 0.023 (250 nM), *p = 0.011 (750 nM), **p = 0.0016 (1500 nM) (two-way ANOVA with Tukey’s multiple comparisons test). b Representative FACS plots of Annexin-V (FITC) and propidium iodide (PE), and c quantification of the percentage of positive apoptotic/necrotic cells after 48 h treatment of myoblasts (DM1 and Ctrl) with 100 nM of A1155463 (Sen) or with vehicle (Ctrl), (n = 4 Ctrl and 7 DM1 cell lines); *p = 0.034 (one-way ANOVA with Sidak’s multiple comparisons test). d Representative micrographs of DM1 myoblasts stained for the senescent marker SA-ß-Gal (green) after 72 h treatment with 100 nM of A1155463 or with vehicle. Scale bar: 300 μm. e Quantification of the number of SA-ß-Gal+ cells expressed as fold change ratio of A1155463-treated (Sen) versus non-treated (Veh) cells (n = 4 Ctrl and 7 DM1 cell lines; average of 1064 cells [142-2873 cells] counted per biological sample). *p = 0.015 (one-way ANOVA with Sidak’s multiple comparisons test). f Quantitative real-time PCR for the senescence markers P16 and P21 on primary myoblasts after 72 h of treatment with vehicle or A1155463 (n = 3 Ctrl and 5 DM1 cell lines, except n = 4 for the p16 expression of DM1-Veh group). *p = 0.042 (two-way ANOVA with Tukey’s multiple comparisons test). g Multiplex Luminex assay of SASP factors showing fold change ratio of A1155463-treated (Sen) versus non-treated (Veh) DM1 myoblasts (n = 4 cell lines, except n = 3 for CCL7 and MMP-1 expression). ***p = 0.0005 (CSF3), *p = 0.018 (CXCL1), ***p = 0.002 (CXCL8), *p = 0.041 (CCL2), **p = 0.009 (MMP-1), *p = 0.027 (MMP-3) (two-tailed multiple unpaired T-tests). Data are expressed as means ± SEM. Source data are provided as a Source Data file.