Figure 1.

Rotenone (ROT) induces reactive oxygen species, mitochondrial depolarization, and chromatin condensation/nuclei fragmentation in Jurkat T leukemia cells. (a) Representative light photomicrography showing positive nitroblue tetrazolium (NBT⁺) stained blue-purple precipitate cells (i.e., formazan: FORMz) as positive O₂ ^∙− generation in Jurkat cells cultured in basal glucose medium containing 11 mM glucose (G11) treated with (50 μM) ROT for 24 h. Inset: untreated cells showing negative NBT staining. (b) Fluorescent photomicrography (ex. 450–490 nm, em. 515 nm) illustrating positive 2′,7′-dichlorofluorescein (DCF, arrows) and negative (arrowheads) stained cells as positive/negative H₂O₂ production from Jurkat T cells cultured in G11 treated with (50 μM) ROT for 24 h. (c) Representative fluorescent photomicrography (ex. 450–490 nm, em. 515 nm) illustrates positive green fluorescent stained cells as DiOC₆(3) high-polarized and low-polarized mitochondria and depolarized mitochondria (asterisk) from treated cells with (50 μM) ROT in G11 for 24 h. (d) Representative (merge image) fluorescent photomicrography showing treated cells with (50 μM ) ROT in G11 for 24 h with condensed chromatin (stage I nuclei morphology) and DNA fragmentation (stage II nuclei morphology) analyzed by either AO/EB ((d′), ex. 450–490 nm, em. 515 nm), Hoechst staining ((d′′), ex. 354 nm, em. 442 nm). (e) Representative histograms showing the percentage of dichlorofluorescein positive (DFC⁺) cells. (f) Percentage of DiOC₆(3) low (DiOC₆(3)⁻) cells exposed to increasing concentration of ROT (0–100 μM), assessed at 24 h. (g) The percentage of positive FORMz, DCF, DiOC₆(3), and AO/EB/Hoechst cells exposed to increasing concentration of ROT (0–100 μM) is expressed as a mean percentage ± S.D. from three independent experiments. Magnification ((a)–(d)) 1000x. Magnification inset (a) 600x.