Figure 4

Both kaempferol and kaempferide suppressed SOD1^G85R-induced superoxide in mitochondria. (A) N2a cells expressing mCherry-SOD1^G85R were treated with 3 µM kaempferol or 15 µM kaempferide for 24 h. Subsequently, MitoSOX® Red was added to the cell culture to a final concentration of 5 μM to the cells and incubated for 30 min at 37 °C. (B) The relative fluorescence intensity was quantified by computerized image analysis with ImageJ. Data are expressed as mean ± S.E.M from three independent experiment, based on the fluorescence intensity of SOD1^WT. (C) Traces show typical spectra of DMPO-OH spin adducts generated from H₂O₂ plus Fe²⁺ in the absence (control) or presence of kaempferol and kaempferide. (D) The amount of •OH was semi-quantitatively measured as the formation of DMPO-OH spin adducts by ESR spectrometry. Data is expressed as the mean ± S.E.M of three determinations, based on control. *p < 0.05; **p < 0.01. ***p < 0.001. Scale bar: 50 µm.