Time-course analysis of A23187-inducible surface Ag expression. HL-60 cells (5 × 105) were cultured in a final volume of 2 mL with or without an optimal dose (180 ng/mL) of A23187 as described in Materials and Methods. Cells were harvested at 20, 48, and 96 hours post addition of A23187 and stained with PE-conjugated mouse-antihuman CD83, B7.1, B7.2, ICAM-1, CD40, CD1a, CD33, or IgG subclass-matched control Ab, and analyzed by FACS as described in Materials and Methods. Results displayed are at 20 hours without A23187 treatment (UNTREATED), 20 hours with A23187 treatment (20H TREATMENT), 48 hours with A23187 treatment (48H TREATMENT), and 96 hours with A23187 treatment (96H TREATMENT). Cultures not treated with A23187 showed an identical immunophenotypic profile at 20, 48, and 96 hours (not shown). Histograms display staining intensity of viable (PI-excluding) cells, comparing control Ab (light lines) to specific Ab (heavy lines).
