BioMed Research International / 2009 / Article / Fig 3

Research Article

Chlamydia trachomatis Alters Iron-Regulatory Protein-1 Binding Capacity and Modulates Cellular Iron Homeostasis in HeLa-229 Cells

Figure 3

Effect of CT infection on the interaction of IRP-IRE: cytoplasmic extracts were prepared from CT-infected HeLa cells treated with DFX and FAC, along with their respective controls, at 24 hpi. Furthermore, the extracts were incubated with 32P-labeled RNA probe, which contained the IRE sequence, in binding buffer. For the supershift assay, the corresponding antibodies (IRP-1 and IRP-2) were added. In parallel experiments, samples were treated with 2-mercaptoethanol) for determination of the maximum binding activity. (a) Binding activity of IRP-1/IRE was attenuated in CT-infected HeLa cells as observed in electrophoretic mobility-shift assay. (b) Inhibition of binding was observed with IRP-1 antibodies in CT-infected HeLa cells. (c) Attenuation of binding activity reverted to normal after treatment with chloramphenicol.

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