Figure 1
Figure 1. Cellular localization and iron-exporting function are not impaired in N144D/T, Y64N, and C326S/T Fpn mutants. (A) Localization of mutants is similar to wt Fpn: HEK293T cells were transiently transfected with human wt Fpn-GFP or with indicated Fpn mutants for 24 hours and imaged by epifluorescence microscopy. (B) Iron export of mutants is similar to wt Fpn: control untransfected HEK293T cells or HEK293T cells transiently transfected with human wt or mutant Fpn-GFP constructs were incubated for 24 hours in the presence of 20 μM FAC. Total cellular protein was isolated, and ferritin content was determined by ELISA. Results are presented as the mean and SD of 3 or 4 separate experiments. Bottom panel illustrates similar expression levels of wt and mutant Fpn-GFP constructs as assessed by Western blotting using anti-GFP antibody. Y64N mutant is presented in a different graph because its ferritin ELISA and GFP Western blotting were performed separately from the analyses of the other mutants.

Cellular localization and iron-exporting function are not impaired in N144D/T, Y64N, and C326S/T Fpn mutants. (A) Localization of mutants is similar to wt Fpn: HEK293T cells were transiently transfected with human wt Fpn-GFP or with indicated Fpn mutants for 24 hours and imaged by epifluorescence microscopy. (B) Iron export of mutants is similar to wt Fpn: control untransfected HEK293T cells or HEK293T cells transiently transfected with human wt or mutant Fpn-GFP constructs were incubated for 24 hours in the presence of 20 μM FAC. Total cellular protein was isolated, and ferritin content was determined by ELISA. Results are presented as the mean and SD of 3 or 4 separate experiments. Bottom panel illustrates similar expression levels of wt and mutant Fpn-GFP constructs as assessed by Western blotting using anti-GFP antibody. Y64N mutant is presented in a different graph because its ferritin ELISA and GFP Western blotting were performed separately from the analyses of the other mutants.

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