Flow cytometric analysis and Western blotting of PSGL-1 and PSGL-1Δcyto transfectants.

(A) HL60 cells (top panels) or K562/FucT-VII cells transfected with either PSGL-1 (middle panels) or PSGL-1Δcyto (bottom panels) were stained with either an isotype-matched negative control (open histograms), the anti-PSGL-1 mAb, KPL1 (left panels, filled histograms), or HECA-452, an antibody that recognizes a reporter epitope associated with FucT-VII activity (right panels, filled histograms). (B) Western blotting of WCLs made from K562/FucT-VII transfectants expressing either PSGL-1 (lane 1) or PSGL-1Δcyto (lane 2). Bands of the appropriate molecular weight were seen in lysates from PSGL-1 (lane 1), and a slight increase in electrophoretic mobility was associated with truncation of 65 amino acids (lane 2).