Fig. 7.

UCH-L1 protects HT-22 cells from H₂O₂ oxidative stress. A and B, HT-22 cells were transfected with empty pcDNA3.1/myc-His(-) A vector (designated as Control) and the same plasmid carrying UCH-L1 WT, C90S, C152S and C90/152S mutant constructs (WT, C90S, C152S and C90/152S, respectively). Cells were treated with 0 or 0.3 mm H₂O₂ in HBSS for 2 h before transferred to RTCA 96-well plate (1 × 10⁴ cells/well) for further analysis. Cell survivals were monitored by xCELLigence real-time cell analysis instrument. C and D, There were no significant differences between 0 mm H₂O₂ treated cells, but wild-type UCH-L1 effectively protected HT-22 cells from 0.3 mm H₂O₂. Cells overexpressing C90/152S mutant were even more vulnerable to oxidative stress. E and F, Overexpressed UCH-L1 with C-terminal myc tag in each experiment was verified by Western blot analysis. Data were shown as the means ± S.E. of triplicates (ns: not significant, *p < 0.05, **p < 0.01).