Fig. 1

SB203580 enhances G-CSF expression in LPS-treated macrophages. RAW264.7 cells (a and b) and BMDMs (c and d) were incubated with DMSO (vehicle) or 10 μM SB203580 for 30 min, then 100 ng/ml of LPS was added and incubation continued for 0 to 16 h, then G-CSF protein levels in the medium were measured by ELISA (a and c) and G-CSF and IL-1β mRNA levels were determined by RT-PCR and analyzed by gel electrophoresis (b and d). (e) RAW264.7 cells were incubated with DMSO or 1, 5, or 10 μM SB203580 for 30 min, then LPS (100 ng/ml) was added and incubation continued for 6 h, then G-CSF and IL-1β mRNA levels were determined by RT-PCR (upper panel). Levels of G-CSF mRNA were determined by RT-qPCR (lower panel), normalized to GAPDH mRNA levels, and expressed relative to the value in the cells treated only with LPS, set as 1. (f) THP-1 macrophages were pretreated with DMSO or 10 μM SB203580 for 30 min, then LPS or PBS was added and incubation continued for 0 to 6 h. Total RNA was then isolated and levels of G-CSF and GAPDH (internal control) mRNA determined by RT-PCR and analyzed by gel electrophoresis. The results in (a) and (e) are the mean ± SD for three independent experiments. *p < 0.05, compared to the group treated only with LPS. (d) The data shown in (b) and (f) are typical of the results obtained in three independent experiments. The data shown in (c) and (d) are typical of the results obtained in two independent experiments