Figure 1

IH increases ROS generation and induces cell death. Aa, b: Images (shown with pseudocolor) and analysis of DCFDA and HE fluorescence staining for ROS. The average fluorescence of the RA4day group was set at 100%. *p = 0.058 and **p < 0.01 for comparing each group with RA4day group by Dunnett's test in ANOVA for DECFA. #p < 0.01 for comparing each group with RA4day group by Dunnett's test in ANOVA for HE. Ba: Cells were costained with Hoechst (blue), TUNEL (green), and Tau (red). Tau was as used as a neuronal marker. Bb: Quantitative assessment of apoptosis based on the area of the nuclei of cerebellar granule cells. **p < 0.01 for comparing each group with RA4day group by Dunnett's test in ANOVA for RA4day and IH1-4 day groups. Bc: Quantitative assessment of apoptosis based on the ratio of TUNEL (+) cells. *p < 0.05 for comparing RA3day with IH3day groups and IH4day by t-test. Ca: Cells were costained with Hoechst (blue), PI (red), and Tau (green). Cb: Quantitative assessment of apoptosis based on the ratio of PI (+) cells. RA4: RA4day group; IH3: IH3day group; IH4: IH4day group. **p < 0.01 for comparing each group with RA4day group by Dunnett's test in ANOVA for RA4day and IH1day-IH4day groups.