Fig. 2.

Self-extracellular RNA/Pam2CSK4 complex promotes pro-inflammatory gene expression in astrocytes in an NF-κB- and MAPK-dependent manner. Astrocytes were treated for 4 h with either (a, b) 5 µM Bay 11–7084, c 20 µM PD98059 or 10 µM SB203580 prior to 8 h stimulation with Pam2CSK4/eRNA molecule complexes, prepared by co-incubation of 100 pg/µl Pam2CSK4 and 1 µg/µl RNA for 2 h at 37 °C. Untreated cells were used as control. a, c Real-time RT-PCR was used to determine transcript levels of pro-inflammatory cytokines. Values are normalized to Rps12 (n = 3–4 per group; One-way ANOVA with Holm–Sidak's multiple comparisons test; * p < 0.05, ** p < 0.01, *** p < 0.001). b TNF-α and IL-6 protein levels in cellular supernatants were quantified by ELISA (n = 3–4 per group; One-way ANOVA with Holm–Sidak's multiple comparisons test; * p < 0.05, ** p < 0.01, *** p < 0.001)