Effects of hybrids on the mitochondria. OP cultures were treated for 24 h with different concentrations (0.5, 1, and 5 µM) of the synthetic compounds, and cell viability was determined by 3-(4,5-dimethyl thiazol-2-y1)-2,5-diphenyl tetrazolium bromide (MTT) assay (A). TMRE fluorescence of single mitochondria were measured after 1-h treatments at different concentrations (0.5, 1, and 5 µM) of compound 1 (comp 1), curcumin (Curc), DMF, or compound 2 (comp 2) and were shown in the histogram (B). TMRE fluorescence of single mitochondria were also measured at different time points (1, 6, 24, and 48 h) during treatments at the chosen concentrations of 1 µM (comp 1 and Curc) and 10 µM (DMF) and shown in the scatter graphs (C). Experimental points are mean ± SEM from n = 80–540 mitochondria. OP cultures were treated for 24 h with 10-ng/mL TNF-α alone or in the presence of comp 1 or Curc. Fluorescence from TMRE-loaded mitochondria was measured and was shown in the bar graph. CTR is taken as 100% (mean ± SEM; n = 60–460 mitochondria; * p < 0.05 vs. CTR) (D). OP cultures treated for 24 h with 10-ng/mL TNF-α were pretreated or not for 24 h with DMF, comp 1, or Curc. After 48-h treatment, the fluorescence from TMRE-loaded mitochondria was measured and was shown in the bar graph. CTR is taken as 100% (mean ± SEM; n = 60–460 mitochondria; * p < 0.05 vs. CTR and # p < 0.05 vs. TNF) (E).