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. 2018 Aug 29;12:622. doi: 10.3389/fnins.2018.00622

FIGURE 1.

FIGURE 1

High-mobility group box 1 (HMGB1) stimulates trophic factor expression in mouse spinal cord astrocytes. Astrocytes from the spinal cord of wild-type mice were pre-incubated for 1 h in the absence or in the presence of the RAGE antagonist FPS-ZM1 (50 nM) or the TLR4 antagonist CLI-095 (5 μM), and then treated with recombinant disulfide-HMGB1 (3 μg/ml) for 6 h. Total RNA was extracted and analyzed by RT-qPCR. Values (mean ± SEM) were normalized relative to hypoxanthine guanine phosphoribosyl transferase (HPRT) and expressed as percentage of untreated cells (ctrl) (main charts). ∗∗∗p < 0.0001 HMGB1 vs. crtl; ###p < 0.0001 HMGB1+FPS-ZM1 or +CLI-095 vs. HMGB1, one-way ANOVA followed by Bonferroni post hoc test, n = 3–17 experiments, in triplicate. Intracellular BDNF and GDNF protein levels from wild-type spinal cord astrocytes were quantified by ELISAs after treatment with recombinant disulfide-HMGB1 (3 μg/ml) for 6 h. Values (mean ± SEM) were expressed as percentage of untreated cells (ctrl) (inset charts). ∗p < 0.05 HMGB1 vs. ctrl, two-tailed unpaired t-test, n = 6 experiments, in duplicate.