Fig. 4

JNK and NF-κB inhibitors suppress the production of pro-inflammatory cytokines induced by LRP1 knockdown. a, b Mouse primary microglia were transiently transfected with non-targeting siRNA (NT) or LRP1-specific siRNA#1 for 48 h. Cells were stimulated with 100 ng/mL LPS or vehicle for 4 h in the presence or absence of 10 μM SP600125 (pretreated for 30 min). The qRT-PCR analysis was then performed to detect the expression levels of IL-1β (a) and TNF-α (b) (n = 3). β-Actin was used as an internal control. c, d Mouse primary microglia were transiently transfected with non-targeting siRNA (NT) or Lrp1-specific siRNA#1 for 48 h. The cells were then pretreated with 10 μM Bay 11-7082 for 30 min, followed by treatment with 100 ng/mL LPS or vehicle for 4 h. RNA was extracted and the relative mRNA levels of IL-1β (c) and TNF-α (d) were determined by qRT-PCR (n = 3). β-Actin was used as an internal control. Data were plotted as mean ± SEM and normalized to the corresponding control group. e, f Reduction of Lrp1 mRNA on siRNA-mediated knockdown was verified by qRT-PCR. *p < 0.05; **p < 0.01; ***p < 0.001; N.S. not significant (one-way ANOVA with post hoc Tukey’s t test)