Figure 3.

Ixazomib reduces nuclear translocation of NF-κB subunit. A, Ixazomib inhibits proteasomal degradation of phosphorylated IκB-α. Raji and Daudi cells were treated with 100 nmol/L concentration of ixazomib for indicated time points. At the end of treatment, total cell lysates were prepared, and immunoblot analyses were performed for pIκBα, t-IκBα, and p65 proteins. The expression levels of β-actin in the lysates served as the loading control. B, Ixazomib reduces nuclear translocation of NF-κB subunit demonstrated by immunostaining. Raji and Daudi cells were treated with 100 nmol/L ixazomib for indicated time points and cytospun onto glass slides. The cells were fixed, permeabilized, and stained for p65 and DAPI. Images were acquired with a fluorescent microscope using a 60× oil immersion lens. C, Ixazomib reduces nuclear translocation of NF-κB subunit demonstrated by cellular fractionation. Raji and Daudi Cells were treated with 100 nmol/L ixazomib for indicated time points, and immunoblot analysis of p65 was performed on subcellular fractions (nuclear and cytosolic). The expression of GAPDH and Lamin B served as the cytosolic and nuclear fraction controls, respectively. Visual overview, Schematic diagram of NF-κB signaling inhibition by ixazomib.