Two-color immunofluorescence analysis on the cultured Lin⁻c-kit⁺ HPCs. Lin⁻c-kit⁺ HPCs from TNF-Rp55^+/+ wild-type or TNF-Rp55^−/− mice bone marrow were cultured in the presence or absence of mouse or human TNFα combined with GM-CSF and SCF for 14 days, and two-color immunfluorescence analysis was performed to determine the expression of Ia with DEC-205 (A) or CD86 (B) as described in the Materials and Methods. Representative results from three independent experiments are shown here.