Subcellular localization of THOC5. A: Schematic representation of subcellular localization and immunostaining of endogenous THOC5. In immature macrophages, THOC5 is located mainly in the nucleus (nuclear speckle-like domains), while in mature macrophages THOC5 is also found in the cytoplasm . Immunostaining: Human mesenchymal stem cells were fixed by methanol and then used for THOC5 specific staining using THOC5 monoclonal antibody . THOC5 is located mainly in the nuclear speckles. B: Nuclear/cytoplasmic shuttling of THOC5 in Hela/mouse 3 T3 cells. Heterokaryon assay: myc-tagged THOC5 was expressed transiently in Hela cells which were then fused with mouse NIH/3 T3 cells in the presence of cycloheximide. Cells were stained by anti Myc-antibody followed by tetramethylrhodamine isothiocyanate (TRITC)-conjugated anti mouse IgG and with 4′,6-diamidino-2-phenylindole (DAPI). *: fused Hela cells; M: mouse NIH /3 T3 cells .