Figure 3

Kaempferol and kaempferide prevented SOD1^G85R-related neurotoxicity. (A) Chemical structures of kaempferol and kaempferide. (B) Representative fluorescent microscopy images of N2a cells expressing mCherry-SOD1^G85R incubated for 24 h with 3 µM kaempferol or 15 µM kaempferide. (C) Quantified data of intracellular SOD1 aggregates in N2a cells expressing mCherry-SOD1^G85R incubated for 24 h with kaempferol (from 0.3 µM to 10 µM) or kaempferide (from 5 µM to 50 µM) are expressed as mean ± S.E.M from three independent experiments. In each experiment, at least 200 cells were counted. (D) Reduction of Triton-insoluble mutant SOD1^G85R by kaempferol and kaempferide. After treatment of 3 µM kaempferol or 15 µM kaempferide, Triton-insoluble fraction of N2a cells expressing SOD1^G85R was resuspended with 2% SDS and analyzed with immunoblotting. (E) The band density of Triton-insoluble mutant SOD1^G85R group is given as mean ± S.E.M from three independent experiments, based on the band density of Triton-insoluble non-treated mutant SOD1^G85R group. (F) N2a cells expressing mCherry-SOD1^G85R were treated with kaempferol (from 0.3 µM to 10 µM) or kaempferide (from 5 µM to 50 µM). The cell viability was measured by MTT assay. Data is expressed as mean ± S.E.M from three independent experiments, based on SOD1^WT group. *p < 0.05; **p < 0.01; ***p < 0.001. Scale bar: 20 µm.