Cytoprotective effect of SB against 6-OHDA damage in SH-SY5Y cells: (A) SH-SY5Y cells were pretreated with 0.1, 1, 10, or 100 nM SB for 1 h and then challenged with 20 μM 6-OHDA for 16 h. Apoptosis in the 6-OHDA-treated group was normalized to 0%. Data are presented as mean ± SEM, and each value represents the mean of three replicates and six samples. * significantly different from the 6-OHDA group; (B) SH-SY5Y cells were pretreated with 0.1 nM SB for 1 h and then challenged with 20 μM 6-OHDA for 8 h. Hoechst 33342 stainings of the control, 6-OHDA, 6-OHDA plus SB, and SB alone groups are shown. The white arrows indicate the locations of chromatin condensation (scale bar = 100 μM); (C) Quantification of cytotoxicity in each group. Data are presented as mean ± SEM, and each value represents the mean of three replicates and three samples. *significantly different from the control group; # significantly different from the 6-OHDA group. p < 0.05

Cytoprotective effect of SB against 6-OHDA damage in SH-SY5Y cells: (A) SH-SY5Y cells were pretreated with 0.1, 1, 10, or 100 nM SB for 1 h and then challenged with 20 μM 6-OHDA for 16 h. Apoptosis in the 6-OHDA-treated group was normalized to 0%. Data are presented as mean ± SEM, and each value represents the mean of three replicates and six samples. * significantly different from the 6-OHDA group; (B) SH-SY5Y cells were pretreated with 0.1 nM SB for 1 h and then challenged with 20 μM 6-OHDA for 8 h. Hoechst 33342 stainings of the control, 6-OHDA, 6-OHDA plus SB, and SB alone groups are shown. The white arrows indicate the locations of chromatin condensation (scale bar = 100 μM); (C) Quantification of cytotoxicity in each group. Data are presented as mean ± SEM, and each value represents the mean of three replicates and three samples. *significantly different from the control group; # significantly different from the 6-OHDA group. p < 0.05

The anti-apoptotic effect of SB on 6-OHDA-induced neurotoxicity in SH-SY5Y cells: SH-SY5Y cells were pretreated with 0.1 nM SB for 1 h and then challenged with 20 μM 6-OHDA for 8 h in the control, 6-OHDA, 6-OHDA plus SB, and SB alone treatment groups. (A) TUNEL staining. White arrows indicate apoptotic cells (scale bar = 100 μM); (B) Quantification of apoptotic cells in each treatment group; (C) Western blotting showing induction of cleaved caspase-3 protein; (D) Quantification of relative density of cleaved caspase-3 protein from Western blotting. Data are presented as mean ± SEM, and each value represents the mean of three replicates and three samples. *significantly different from the control group; # significantly different from the 6-OHDA group.

Effect of SB on 6-OHDA-induced upregulation of intracellular reactive oxygen species (ROS) and downregulation of superoxide dismutase (SOD) activity. SH-SY5Y cells were pretreated with 0.1 nM SB for 1 h and then challenged with 20 μM 6-OHDA for 2 h. (A) CellROX^® of the control, 6-OHDA, 6-OHDA plus SB, and SB treatment groups. White arrows indicate the location of ROS-positive cells (scale bar = 100 μM). (B) Quantification of the frequency of ROS-positive cells in each group. (C) SOD levels of the control, 6-OHDA, 6-OHDA plus SB, and SB alone treatment groups. Data are presented as mean ± SEM, and each value represents the mean of three replicates and three samples. *significant compared with the control group; # significant compared with the 6-OHDA group.

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