Fig. 3: Electron transfer chain perturbation by NCA. | Cell Death Discovery

Fig. 3: Electron transfer chain perturbation by NCA.

From: Neocarzilin A induces apoptosis and mitochondrial disturbance by targeting reticulon 4-mediated endoplasmic reticulum stress

Fig. 3

A Oxygen consumption rate (OCR) of HeLa cells treated with NCA, DMSO, and antimycin A as positive control was assessed in a plate reader assay over 6 h at 37 °C using an oxygen-quenchable fluorescent dye. OCR was calculated from the slope of the recorded curves. B High-resolution respirometry measurements of 6 h NCA or DMSO-treated cells following SUIT protocol 008_O2_ce-pce_D025 in an Oroboros O2k instrument. C Complex I activity of mitochondria enriched from HeLa cells treated with NCA or DMSO as indicated was measured by NADH-dependent indirect reduction of DCIP using decylubiquinone as substrate. Decrease in absorbance at 595 nm was recorded over 15 min in a plate reader, not complex I-related conversion was assessed by applying rotenone. Enzymatic activity was calculated from the slope of the linear section of the curves and the protein amount. D CellTiter-Glo assay to determine ATP content of NCA or DMSO stimulated HeLa cells as indicated. Glycolysis was inhibited by concomitant 2-deoxyglucose (200 mM, 6 h) treatment. C Absolute values, A, D normalized data and C control efficiencies are presented in bar graphs as mean ± SD, A, D n = 3, C n = 4, B n = 5. Statistical significance was analyzed by B, C two-tailed unpaired Student’s t-test or Mann–Whitney test or A, D one-way ANOVA with Dunnett’s posttest compared to mean of DMSO control (*P < 0.05, **P < 0.01, ***P < 0.001).

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