PBMCs were stained for surface markers CD4 and CADM1 following LIVE/DEAD cell staining (Invitrogen, L34976). Then the viral protein Tax was stained intracellularly with Foxp3 staining kit (eBioscience, 00-5523-00) (
Figure 2a ). CADM1 staining was included in order to obtain an equivalent number of HTLV-1-infected cells in the Tax
– population
20 (link). HTLV-1-infected T cell clones were stained with LIVE/DEAD and anti-Tax antibody (
Figure 2b ). The antibodies used were: mouse anti-CD4-PE (clone RPA-T4; BioLegend, 300507; concentration used, 0.8 μg/ml); chicken anti-CADM1-biotin (clone 3E1; MBL, CM004-6; 20 μg/ml) in combination with SA-BV421 (BioLegend, 405226; 1 μg/ml); mouse anti-Tax-Cy5 or anti-Tax-AF647 (clone LT-4; 0.4 μg/ml)
21 (link). Cell sorting was carried out with a BD FACSAria III.
– population
20 (link). HTLV-1-infected T cell clones were stained with LIVE/DEAD and anti-Tax antibody (
21 (link). Cell sorting was carried out with a BD FACSAria III.
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