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Fig. 3 | Journal of Neuroinflammation

Fig. 3

From: 1-Oleyl-lysophosphatidic acid (LPA) promotes polarization of BV-2 and primary murine microglia towards an M1-like phenotype

Fig. 3

LPA promotes classical activation of BV-2 and primary microglia. a Serum-starved BV-2 cells were treated with BSA (0.1 %; ‘c.’), LPA (1 μM), LPS (20 ng/ml), IL-4 (40 ng/ml), or IL-10 (40 ng/ml)], and cellular protein lysates were analysed by Western blotting. LPS, IL-4, and IL-10 were used to polarize cells to an M1- or M2-like phenotype, respectively. One representative plot for each protein and the densitometric analysis (mean + SD; normalized to actin) from four independent experiments is presented. Control = 0.1 % BSA. b Confocal immunofluorescence microscopy of PMM in the absence or presence of LPA (1 μM, 24 h). LPS (20 ng/ml) and IL-4 (40 ng/ml) were used to induce an M1- or M2-like phenotype, respectively. Cells were stained for CD11b (microglia marker) and COX-2 or Arg-1. Nuclei were counterstained with Hoechst. Scale bars = 20 μm. Results from one representative experiment (out of two) are shown. c PMM cultured on chamber slides were incubated in the absence or presence of LPA (1 μM) for 24 and 48 h. Cells were stained for specific inflammatory markers and nuclei were counterstained with Hoechst. The fluorescence intensity for each marker was quantitated with ImageJ. At least 50 cells out of 3 different areas per chamber were measured in two independent experiments. The results are presented as mean + SD (*p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA with Bonferroni correction)

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