Fig. 7.
Failure of C2-ceramide to inhibit p38 MAP kinase phosphorylation.
(A) PMNs were treated with 10 μM C2-ceramide or buffer for 30 minutes at 22°C. Cells were then stimulated with 50 ng/mL G-CSF for 10 minutes at 37°C, alone or followed by 1 μM FMLP for 1 or 3 minutes. Cells were lysed with Triton X-100 and samples run on 10% SDS-PAGE. Western blots were probed with antibody against phosphorylated p38 MAP kinase. One representative experiment of 3. (B) The Western blot from panel A was stripped and reprobed with anti-p38 MAP kinase that recognizes phosphorylated and nonphosphorylated forms to show equal loading.
