Figure 5.

Cytoplasmic HBZ is retained in this compartment and does not shuttle to the nucleus. (A) Peripheral blood mononuclear cells (PBMC) of representative acute PH140126 and chronic PH170918, were either treated (+LMB, bottom panels) or not treated (-LMB, top panels) with Leptomycin B (LMB), an inhibitor CRM1/exportin-mediated nuclear export, before fixing and stained with the anti-HBZ 4D4-F3 monoclonal antibody (mAb) followed by Alexa Fluor 546-conjugated goat anti-mouse IgG1 antibody (red) and analyzed by confocal microscopy. (B) As control of inhibition of nuclear export by LMB, treated (+LMB) and untreated (-LMB) cells were fixed and stained with antibodies against p65/RelA followed by Alexa Fluor 546-conjugated goat-anti-rabbit secondary antibody (red) and analyzed by confocal microscopy. One representative image on the field and an enlargement of this are shown. Nucleus was stained with DRAQ5 (blue). DIC represents the differential interference contrast image. One representative image is shown for each sample. At least 300 cells were analyzed. All scale bars are 5mm.