Fig. 3

RAP increases the expression of pro-inflammatory cytokines and induces both JNK and NF-κB activation in microglia. a–c Primary microglia were treated with control, LPS (100 ng/ml) or RAP (25 or 50 nM) for 4 h (for qRT-PCR) or 30 min (for Western blotting). The qRT-PCR was performed to determine mRNA levels for Il-1β (a), Tnf-α (b), and Lrp1 (c) (n = 3). d–h The protein levels of phospho-IκBα, total IκBα, phospho-NF-κB, total NF-κB, phospho-c-Jun, total c-Jun, and LRP1 in cell lysates were examined by Western blot analysis (d) and quantified (e–h) (n = 3). β-Actin served as a loading control. Data were plotted as mean ± SEM and normalized to the corresponding control group. *p < 0.05; **p < 0.01; ***p < 0.001; N.S. not significant (one-way ANOVA with post hoc Tukey’s t test)