Figure 3: Efficiency of the MCU blocker Ru265 in LS8 enamel cells.

A-B) Quantification of mitochondrial Ca²⁺ uptake in rhod2AM (4 μM) loaded LS8 cells (~120 cells/field per condition) treated with Ru265 for 1 h at the following concentrations: 1, 10, 20 and 50 μM. Data were analyzed by one-way ANOVA. ****P < 0.0001 C) LogEC₅₀ plot for data in A and B. D) Quantification of Ca²⁺ clearance in digitonin-permeabilized LS8 cells (n = 800 K cells) loaded with Calcium Green-5 N (1 μM) in the presence of Ru265 (5 μM) or RuR (5 μM) after the application of Ca²⁺ boluses (20 μM). E) Mitochondrial membrane potential (ΔΨm) measured using TMRM (40 nM) in LS8 cells (n = 200 K). Oligomycin A (5 μM) and FCCP (5 μM) were added to the cells to induce mitochondria hyper- and depolarization, respectively. Data represent mean ± SEM, from a minimum of 3 independent experiments.