Subcellular localization of HBZ and THEMIS.
(A) Staining of unstimulated or pervanadate-stimulated Jurkat-mock and Jurkat-HBZ cells was performed using antibodies against PD-1 (green) and THEMIS (red). All scale bars are 5 μm. Three representative images derived from each sample are shown. Relative fluorescence intensities of PD-1 (green line) and THEMIS (red line) were obtained over white dotted line. (B) Localizations of HBZ (red) and THEMIS (green) were analyzed in HBZ and/or THEMIS-transfected 293T cells. Nucleus was stained with DAPI (blue). Co-localization of HBZ and THEMIS was found in co-transfected cells (indicated by arrowheads). Two representative images derived from each sample are shown. All scale bars are 30 μm. (C) Localization of HBZ in Jurkat-HBZ cells was detected using anti-HBZ antibody. Nucleus was stained with DAPI (blue). Scale bar is 10 μm. (D) Expression of THEMIS was detected by immunoblotting in 293T cells and Jurkat cells. (E) Effect of THEMIS on the localization of HBZ was analyzed in myc-tagged HBZ-introduced THEMIS-KD and luciferase-KD (served as a control) Jurkat cells. shRNA-expressing cells express GFP as a marker (shown in gray). Cells were stained with antibodies against nuclear pore complex (NPC; green) and myc-tag (HBZ; red). Five representative images derived from each sample are shown. All scale bars are 5 μm.
