American Society of Hematology

Fig. 4.

$Fig. 4. Failure of C2-ceramide to inhibit mobilization of secretory vesicles. / PMNs were treated with 10 μM C2-ceramide (C2), then activated with 5 μg/mL cytochalasin B (CB) and 100 nM FMLP (F) (triangles) as described in Figure 3. Controls were unstimulated (closed circles) or activated in the same way without C2-ceramide (open circles) and were incubated in parallel. Samples were N2-cavitated, loaded on a 3-layer Percoll gradient, and fractionated. Alkaline phosphatase activity was determined in the presence and absence of Triton X-100, and gelatinase by ELISA. (A). Latent alkaline phosphatase in fractions representing secretory vesicles and plasma membrane. (B) Gelatinase in tertiary granule fractions. Data represent the mean ± SEM of 6 experiments. *C2-ceramide–treated sample (C2 + CB + F) significantly different from stimulated control (CB + F); P < .05.$

Failure of C₂-ceramide to inhibit mobilization of secretory vesicles.

PMNs were treated with 10 μM C₂-ceramide (C2), then activated with 5 μg/mL cytochalasin B (CB) and 100 nM FMLP (F) (triangles) as described in Figure 3. Controls were unstimulated (closed circles) or activated in the same way without C₂-ceramide (open circles) and were incubated in parallel. Samples were N₂-cavitated, loaded on a 3-layer Percoll gradient, and fractionated. Alkaline phosphatase activity was determined in the presence and absence of Triton X-100, and gelatinase by ELISA. (A). Latent alkaline phosphatase in fractions representing secretory vesicles and plasma membrane. (B) Gelatinase in tertiary granule fractions. Data represent the mean ± SEM of 6 experiments. *C₂-ceramide–treated sample (C2 + CB + F) significantly different from stimulated control (CB + F); P < .05.

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