Fig. 6.
Effects of JNK and p38 MAPK on PU.1-driven transcription and L-PGDS expression in macrophages. RAW 264.7 cells were transfected with pCMV (columns 1 and 2) or a plasmid encoding PU.1 (columns 3–6) or PU.1S148A (columns 7–10), along with tk-Renilla luciferase construct and a PU.1 firefly luciferase reporter construct. At 48 h after transfection, the cells were treated with either JNK inhibitor (columns 5 and 9) or p38 kinase inhibitor (columns 6 and 10) 1 h before LPS treatment for 4 h. Luciferase activity was measured from triplicate of the experimental settings, and the experiment was performed 3 times independently. Each bar represents the mean RLU ± SE. *P < 0.05 compared with the control value (lane 1). ** and ***P < 0.05 in comparison of lane 4 with lanes 5 and 6, respectively. B: similar experiment was performed with a PU.1-luciferase reporter that contains 3 repeats of the PU.1 binding site of the murine L-PGDS promoter. C: RAW 264.7 cells were treated with vehicle (lanes 1 and 2), 10 μM JNK inhibitor (lane 3), or 10 μM p38 kinase inhibitor (lane 4) 1 h before LPS treatment for 16 h to induce L-PGDS. Western blot was performed for L-PGDS and tubulin.