Figure 8. KP46 sensitizes the mitochondrial PTP to Ca²⁺.

a. CRC was performed on HCT116^WT cells exposed to 2.5 μM KP46 (grey trace) or DMSO (black trace) for 4 h. b–c. Thapsigargin partly prevents Ca²⁺ induced PTP opening of KP46 treated cells. CRC experiments on HCT116^WT cells exposed to 2.5 μM KP46 (b) or vehicle (c), co-treated with CsA (10 μM) added prior measuring or NAC (1 mM) added 1 hour prior and during the KP46 treatment or Thapsigargin (1 μM) added during KP46 treatment. d. Quantification of HCT116^WT cells treated with 2.5 μM KP46 or DMSO for 4 h with or without the addition of Thapsigargin, CsA or NAC (n = 3, if not mentioned separately) ±SEM, ****p< 0.0001, **p < 0.01, two-way ANOVA, followed by Bonferroni's multiple comparisons test. e. BNIP3L knockdown prevents PTP opening of KP46 treated cells. CRC were performed as in (a) on HCT116^WT transfected with shRNAscr or shRNABNIP3L and treated with 2.5 μM KP46. f. Quantification of HCT116^WT cells scramble transfected or with Bnip3L KD were treated with KP46 2.5 μM or vehicle for 4 h. n = 3, ±SEM, ****p < 0.0001, ***p < 0.001, **p < 0.01, two-way ANOVA, followed by Bonferroni's multiple comparisons test.