Figure 3.

Deferasirox inhibits NF-κB activity in MDS patients. (A) NF-kB DNA binding activity (EMSA assay) in a MDS patient: NF-κB activity can be detected in control cells and its amount decreases after 18 h of deferasirox incubation in vitro at 50 mM. The black arrow indicates specific binding to the consensus sequence. The second lane of each condition was loaded with an unspecific cold probe as negative control. (B) Inhibition of NF-κB activity after in vitro incubation with 50 mM deferasirox of mononuclear PB cells of MDS patients for 18 h, as measured by ELISA assay. In all patients with high basal NF-κB activity (28 patients out of 40 tested), it is possible to detect transcription factor (TF) inhibition after drug incubation. (C) Immunofluorescence assay of PB cells from an MDS patient. At basal conditions on the left, p65 is completely localized in the nucleus, but after deferasirox incubation, the subunit is largely located in the cytoplasm, as demonstrated by the presence of the green signal.