Fig. 2.

Ndfip1 protects human neurons from metal toxicity. (A) Tamoxifen-inducible expression construct of Ndfip1-Flag was created and used in a stable line of SH-SY5Y cells (anti-Flag blot shown). (B) Flow cytometry profile of SH-SY5Y cells treated with Co for 18 h. Different distributions of alive (quadrant 1), apoptotic (quadrant 2), and apoptotic plus necrotic populations (quadrant 3) between control and treated cells are shown. Cells treated with Co are dying through an apoptotic pathway. (C) Flow cytometry analysis of SH-SY5Y cells induced to express exogenous Ndfip1 protein protects against Co toxicity, compared with uninduced controls. (D) Flow cytometry analysis indicates that human embryonic cortical neurons induced to express exogenous Ndfip1 are protected against Co toxicity, compared to uninduced controls. (E) SH-SY5Y cells stained for Fe uptake show that cells induced to express Ndfip1 prevented Fe uptake compared with uninduced controls. Two-way Anova tests (Bonferroni posttests) indicate significant protection by Ndfip1 at higher concentrations of Co in (C) and (D) (P < 0.05* and P < 0.001***, ± SEM). Each flow cytometric analysis represents three independent experiments.