Effect of TMQ0153 on cellular metabolism.a Oxygen consumption rate (OCR) was measured by Seahorse XFp analyzer. b Caspase 8 analysis by western blot (upper panel) and the corresponding densitometric analysis (lower panel). Etoposide (Eto; 100 µM, 24 h) was used as a positive control for apoptotic caspase cleavage. c Caspase 8 analysis by western blot (upper panel) in the presence or absence of autophagy inhibitors: 40 nM baf-A1, 10 mM, 3-methyladenine (3-MA) and 75 μM chloroquine (CQ) and the corresponding densitometric analysis (lower panel). In western blot analyses, β-actin was used as a loading control. d Quantification of caspases-3/7 (left graph) and -9 activity (right graph) levels at 30 µM of TMQ0153 treatment. Etoposide (Eto; 100 µM, 24 h) was used as a positive control for apoptosis induction. All pictures are representative of three independent experiments and data represent mean (±S.D.) of three independent experiments. Statistical significance was assessed as *p < 0.05, **p < 0.01, ***p < 0.001 compared to untreated cells unless otherwise specified. One-way ANOVA (Caspase-3/7 and -9 assay); post hoc; Tukey’s test. Two-way ANOVA (mito stress test); post hoc; Sidak’s test. One-way ANOVA (western blot quantification); post hoc; Dunnett’s test.
|