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. Author manuscript; available in PMC: 2011 Sep 17.
Published in final edited form as: Cell. 2010 Sep 17;142(6):857–867. doi: 10.1016/j.cell.2010.08.014

Figure 4. Intracellular iron accumulates in APP−/− neurons.

Figure 4

A, APP−/−; primary neurons treated with Tf(59Fe)2 retain more 59Fe after 12 h than cells from WT controls. APP695α (2 μM) promotes 59Fe export into the media after 12 h from both WT and APP−/−; neurons. In APP−/−; neurons this reduces intracellular iron to approach WT levels. B, 59Fe media efflux is decreased for APP−/−; compared to WT primary neurons. Data are 59Fe counts in media expressed as a fraction of the total in culture. C, Western blot (see Figure S3D) quantification of ferritin and TfR in primary neuronal cultures from WT and APP−/−; matched controls treated ± Fe(NH4)2(SO4)2 (75 μM). Differences in APP−/−; cells are consistent with increased retention of iron. D, APP and Cp co-immunoprecipitate with ferroportin from human and mouse brain, but not APLP2. E, Determination that membrane-bound full-length APP interacts with ferroportin using APP detection antibodies for both the N- and C-terminal ends of the protein from membrane lysate of human brain immunoprecipitated by anti-Fpn antibody. F, APP−/−; neurons incubated with increasing concentrations of Fe(NH4)2(SO4)2 are more susceptible to iron toxicity, measured by CCK-8 cell viability assay, than WT neurons. Data are means ± SEM, n=3, *= p<0.05, **= p<0.01, ***= p<0.001, A - C analysed by 2-tailed t- tests, D by ANOVA + Dunnet’s test compared to WT. See also Figures S3 and S4.