TRANCE and RANK are expressed by CD34⁺-derived int-DCs.

(A) CD34⁺-derived int-DCs, unstimulated or CD40L-stimulated DCs, generated in the presence of GM-CSF and TNFα25 and anti–CD3/CD28-activated T cells were tested for the transcription of TRANCE, RANK, OPG, and β-actin by RT-PCR. (B) COBS cells were transfected with Flag-tagged human TRANCE cDNA, and cDNA expression was analyzed by anti-Flag and anti-TRANCE Ab. Percentage of positive cells is indicated. Detection of RANK (C) and TRANCE (D) by Western blot in the indicated cells. Full-length RANK and recombinant soluble RANK-Fc migrated with an apparent molecular weight of about 90 kDa and about 70 kDa, respectively, and full-length TRANCE and recombinant soluble TRANCE with an apparent molecular weight of about 42 kDa and about 20 kDa, respectively. CD1a⁺ L-DCs and macrophages served as positive controls; Hela cells and the myeloma U266 served as negative controls.