Expression of hCLEC9A on human blood DCs. (A) Human peripheral blood mononuclear cells (PBMCs) were isolated and surface immunofluorescence labeled with mAb against HLADR (L243-PercP), a cocktail of PE-conjugated mAb against Lineage markers including CD3 (BW264156; T cells), CD14 (Tuk4; monocytes), CD19 (6D5; B cells) and CD56 (AF12-7H3; NK cells), hCLEC9A (3A4-FITC), and either CD16 (VEP-APC-Cy7) or BDCA-1 (ADJ-8E7-APC), or BDCA-3 (AD5-14H12-APC), or BDCA-4 (AD5-17F6-APC; pDC). Blood DCs were gated as HLADR⁺Lineage⁻ and further analyzed for their expression of hCLEC9A. (B) Other heamapoeitic cells were identified in preparations of PBMCs by immunofluorescence labeling with mAb against CD19 (6D5-PE; B cells), CD3 (BW264156-PE; T cells), CD56 (AF12-7H3-PE) and NKp46 (9E2-APC; CD56⁺NKp46⁺; NK cells), CD14 (Tuk4-PE; monocytes), together with 3A4-FITC, then analyzed for their expression of hCLEC9A. The dotted line represents the background staining of the gated cells with an isotype-matched control.