Figure 1

Acquisition of granulocytic features following differentiation of PLB-985 cells. (A) Cells were differentiated for 5 days in the presence of DMF or DMSO, or left undifferentiated, prior to FACS determination of their size (forward scatter) and granularity (side-scatter). (B) Cells were differentiated for 5 days in the presence of DMF or DMSO, or left undifferentiated, prior to staining with an anti-CD11b Ab (black trace), with an isotype-matched control Ab (grey trace) or without a first Ab (dotted trace) and subsequent incubation with a FITC-conjugated 2nd Ab. CD11b surface expression was then determined by FACS analysis; a minimum of 10,000 cells were processed for each sample. Results are representative of at least five independent experiments. (C) Results obtained as described above were compiled and expressed as mean ± s.e.m. of at least 5 independent experiments. For comparison, 100% of human neutrophils express CD11b on their surface. (D) Cells were differentiated for 5 days in the presence of DMF or DMSO, or left undifferentiated, prior to stimulation for 6 h at 37°C with fMLP or LPS. Culture supernatants were then collected and analyzed by ELISA. Mean ± s.e.m. of at least 6 independent experiments.