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. 2015 Aug 24;112(36):E5078–E5087. doi: 10.1073/pnas.1514486112

Fig. S3.

Fig. S3.

IL-1βlow sensitizes its own response via IL-1R1. (A) Experimental design: Primary hippocampal neurons at 5–7 DIV were preincubated with IL-1βlow (3 pM) or vehicle for 3 h. After washing, vehicle-treated neurons were stimulated with vehicle or BDNF only (50 ng/mL, 1 h), whereas neurons pretreated (pre-Tx) with IL-1βlow were stimulated with BDNF in the presence of IL-1βlow for 1 h. To specifically test the role of IL-1R1 on the induction of sensitivity, IL-1ra (300 nM) was added 30 min before IL-1βlow pretreatment, but was absent during stimulation. Control groups consisted of a vehicle-only group (vehicle) and an IL-1ra–vehicle group (IL-1ra). (B) Phosphorylated and total levels of Akt and CREB were evaluated by using Western blots. (C and D) Relative levels of p-Akt(Ser-473)/Akt (C) and p-CREB(Ser-33)/CREB (D) were normalized with vehicle-only treated controls [n = 5 per group; five independent experiments (neurons from different embryo litters)]. *P < 0.05; (*)P < 0.01; [*]P < 0.001 vs. vehicle control (ANOVA, Tukey’s post hoc test). Data are presented as mean ± SEM.