Table A.1.

Quantitative evaluation of the 1st KER: Binding of inhibitor to NADH‐ubiquinone oxidoreductase (MIE; KE upstream) leads to its inhibition (KE downstream)

MIE (KE upstream) Binding of inhibitor to NADH‐ubiquinone oxidoreductase (nM)	KE (downstream) Inhibition of CI (%, approximately)	Comments (in vivo, in vitro or human studies)	References
Administration of rotenone at 2 mg/kg per day for 2 days resulted in free rotenone concentration of 20–30 nM in the brain	75%	DA neuronal cell death determined after rotenone administration at 1–12 mg/kg per day, Sprague–Dawley and Lewis rats infused continuously by jugular vein, 7 days up to 5 weeks	Betarbet et al. (2000)
20 nM rotenone Direct binding studies using bovine and Musca domestica submitochondrial particles	50%	Binding studies that defined the I50 and K d values for three classes of CI inhibitors (12 chemicals) including rotenone	Okun et al. (1999)
Human skin fibroblasts exposed to 100 nM Rotenone for 72 h	20%	In the same experiment mitochondria morphology, motility was also evaluated	Koopman et al. (2007)
0–2.5 nM Rotenone 5/10 nM Rotenone Mesencephalic neurons were cultured from E14 C57BL/6 mouse embryos for 6 days and then treated with rotenone for 24 h	No effect 11% and 33%, respectively	Treatments with 5 or 10 nM rotenone killed 50% or 75% DA neurons respectively	Choi et al. (2008)
1–2.5–5–7.5–10–20 nM 1–10–20–80 nM	10–20–35–50–65–80% 5–75%	In this study time course of the active and deactivated enzymes inhibition by rotenone and piericidin A is study in a dose‐dependent manner Binding studies in submitochondrial particles prepared from bovine heart after 20 min of exposure to rotenone	Grivennikova et al. (1997)
5–10 nM 20 nM 40 nM 100 nM 143B Cells (human osteosarcoma), exposed for 4 h to rotenone	55–78% 80% 87% 100%	In the same study similar experiments were performed using HXC cell line (see Figure A.1 above)	Barrientos and Moraes (1999)