Figure 1.

Inverse correlation between the level of lcn2 expression and the sensitivity of astrocytes to cytotoxic agents. C6 glial cells with an increased or decreased lcn2 expression were established by stable transfection of lcn2 sense (S3) or antisense (AS7) cDNA. The increased or decreased lcn2 expression in the stable transfectants compared with cells transfected with an empty vector (V2) was confirmed by RT-PCR (top) and Western blot analysis (bottom) (A). β-Actin or α-tubulin was also detected to confirm the equal loading of the samples. Compared with the empty vector transfectant (V2), the lcn2 sense (S3) or antisense (AS7) stable transfectants showed an increased or decreased sensitivity, respectively, to cytotoxic agents such as SNP, H₂O₂, and paraquat (B–D). Cells were treated with the indicated concentrations of cytotoxic agents for 24 h, and cell viability was assessed by MTT assay. The asterisks indicate statistically significant differences from the empty vector transfectant (V2) treated with the same concentrations of cytotoxic agents (V2 vs S3 or V2 vs AS7) (p < 0.05). The results are mean ± SD (n = 3). The results in this and all similar experiments were repeated several times, and one representative done in triplicate is shown.