TruCOUNT analysis of absolute DC numbers in cultured PBMCs.

Dot plots demonstrate the forward and side scatter profiles of cultured PBMCs (A, left column). PBMCs were gated in R1, TruCOUNT beads in R2 (A, left column), and Lin⁻ HLA-DR⁺ DCs in R3 (A, right column). Absolute DC numbers were calculated as the mean of triplicate determined by the number of Lin⁻HLA-DR⁺ events per 10 000 TruCOUNT beads acquired for each time point (n = 8, B, left). The filled and hatched portions of the bars represent the proportion of Lin⁻ HLA-DR^loand Lin⁻ HLA-DR^hi cells in R3, respectively. The proportion of Lin⁻ HLA-DR^lo cells is shown as percentage of total DCs. PBMCs were irradiated (3000 Gy) before culture (n = 3, B, right). Error bars show SEM. (**,P < .001).