Fig. 4

MAPK family members differentially regulate the LPA-induced pro-inflammatory phenotype of microglia. a Serum-starved primary microglia were treated with DMSO and DMSO plus LPA (1 μM) in the absence or presence of SP600125 (10 μM), SB203580 (10 μM), or PD98059 (10 μM) for the indicated times. Cell lysates were collected, and protein expression of COX-2 and Arg1 was monitored by immunoblotting. One representative blot for each protein and b the densitometric analysis (mean + SD) from four independent experiments is presented. c–e Serum-starved (overnight) primary microglia were cultivated in the presence of DMSO and DMSO plus LPA (1 μM) in the absence or presence of SP600125 (10 μM), SB203580 (10 μM), or PD98059 (10 μM) for the indicated time periods. Cells were stained with PE-conjugated anti-CD40, APC-conjugated anti-CD86, or PE-conjugated anti-CD206 antibodies and analyzed using a Guava easyCyte 8 Millipore flow cytometer. Results from three individual experiments performed in duplicates are shown as mean values ± SD. (*p < 0.05, **p < 0.01, ***p < 0.001 compared to DMSO-treated cells; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 each individual inhibitor compared to LPA-treated cells; two-way ANOVA with Bonferroni correction)