BEAS-2B cells were stained with Fluo-3 AM (green) and Hoechst 33342 (blue) and analyzed immediately upon treatment with MBP-118-45 in the presence or absence of mtDNA using live-cell confocal microscopy. Ionomycin was used as a positive control. Left: quantification of the calcium levels was performed by automated analysis of microscopy images using Imaris software. Significances are shown against MBP-118-45 alone (black). Right: bars, 20 μm. Data are representative of 3 independent biological replicates. The assay was performed in duplicates. All data are represented as mean ± SEM, and p values (*p< 0.05,**p< 0.01,***p< 0.001) were calculated using ordinary one-way ANOVA with Tukey’s multiple comparisons (A) and two-way ANOVA with Dunnett’s multiple comparisons (B).
