Fig. 6.
Fig. 6. Gel shift assay of IRE-IRP interactions in livers from. / Fth+/+ andFth+/− mice. ThreeFth+/+ (1-3) and 3Fth+/− (4-6) mice were killed and cytoplasmic extracts were prepared from their livers. Equal amounts of proteins were analyzed for IRE binding in the presence or absence of 2% βME, by means of a sense human H ferritin IRE probe. Extracts from RR4, a mouse microglial cell line, were used to control for the position of IRE-IRP1 and IRE-IRP2 complexes. Radioactivity associated with the IRE-IRP1 complex was quantified by means of an Instantimager. IRP1 activity was expressed as a percentage of the value obtained after 2% βME treatment of the cytoplasmic extracts, which allows one to estimate the total IRE bonding capacity of IRP1.

Gel shift assay of IRE-IRP interactions in livers from

Fth+/+ andFth+/− mice. ThreeFth+/+ (1-3) and 3Fth+/− (4-6) mice were killed and cytoplasmic extracts were prepared from their livers. Equal amounts of proteins were analyzed for IRE binding in the presence or absence of 2% βME, by means of a sense human H ferritin IRE probe. Extracts from RR4, a mouse microglial cell line, were used to control for the position of IRE-IRP1 and IRE-IRP2 complexes. Radioactivity associated with the IRE-IRP1 complex was quantified by means of an Instantimager. IRP1 activity was expressed as a percentage of the value obtained after 2% βME treatment of the cytoplasmic extracts, which allows one to estimate the total IRE bonding capacity of IRP1.

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