Effects of MCSF on the phosphorylation of MAPKs.

<p>After pre-incubation for 3 days with 10 ng/ml MCSF, peritoneal macrophages were incubated for 5 and 15 min with 10 ng/ml MCSF. Cellular protein was extracted and protein samples (0.4 mg/ml) were analyzed by Western blot with specific antibodies: anti-phosphorylated MEK1/2 or anti-MEK1/2 (<b>A,</b> right), anti-phosphorylated ERK1/2 or anti-ERK1/2 (<b>B,</b> right), anti-phosphorylated p90RSK or anti-RSK1/2/3 (<b>C</b>, right), anti-phosphorylated p38 MAPK or anti-p38 MAPK (<b>D,</b> right) and anti-phosphorylated SAPK/JNK or anti-SAPK/JNK (<b>E</b>, right) antibodies (representative experiments). ß-Actin or Actin were used as control. Quantitative results were calculated by band densitometry with the intensity of phosphorylated MEK1/2, ERK1/2, p90RSK, p38 MAPK and SAPK/JNK normalized to the total MEK1/2, ERK1/2, RSK1/2/3, p38 MAPK and SAPK/JNK (<b>A–E</b>, left panels). Data represent mean ± SD of 3 separate experiments. *, **indicate statistically significant differences (*p<0.05, **p<0.01) compared with cells without MCSF treatment.