Subcellular distribution of leukemia-type Helios isoforms.

<p>(<b>A, B</b>) Cos-7 cells were transfected with the indicated five Helios isoforms and subjected to RT-PCR (<b>A</b>) and Western blot analyses (<b>B</b>). The COS-7 cell extracts were developed with the mouse monoclonal anti-Flag antibody (<b>left</b>) and an anti-Helios antibody (<b>right</b>). (<b>C</b>) Immunofluorescence localization of Helios proteins. The COS-7 cells were transfected with different Helios isoforms. The cellular localization of each protein was visualized with the anti-Flag antibody and then stained with a secondary anti-mouse antibody conjugated with Cy2 (green). The nuclei were counterstained by DAPI staining (blue). Helios-V1, Helios variant 1; Helios-V2, Helios variant 2; Helios-V3, Helios variant 3. (<b>D</b>) Graphic representation of the subcellular localization of Helios alternative-splicing variants. The analysis was conducted by counting nuclear or cytoplasmic staining, or both (nuclear and cytoplasmic), of the five isoforms as percent point. Eight fields were counted for each transfection.