Fig. 5.

Reversal of α-syn oligomer-induction of cell priming. The pro-inflammatory response (induction of TNF-α) by BV2 microglia after priming with oligomeric α-syn was determined by ELISA. a BV2 microglia were primed with 2 nM, 1 nM or 200 pM concentrations of α-syn oligomers for 72 h before α-syn was removed and replaced with fresh buffer. TNF-α levels were measured every 12 h by replacing the solution above the cells with fresh buffer. The production of TNF-α decreases slowly with time, but is not completely restored to the unprimed state (n = 3, sem). 2 nM, 1 nM and 200 pM oligomer is significantly higher than untreated cells at all times (p = 0029, p = 0.0056 and p = 0.087 respectively). b–d BV2 microglia were incubated with oligomeric α-syn, 2–2000 nM. Cells were primed for different lengths of time (b 24-h, c 48-h, d 72-h) before α-syn oligomers were removed, cells were washed with media and replaced with media containing α-syn oligomers and the TLR4 antagonist RSLA. RSLA reduced TNF-α production and enhanced cell survival returning viability to 90% (± 3%) over the duration of the experiment (Supplementary Fig. 5b) (n = 4, sem). All statistical comparisons among groups were performed using one-way ANOVA with post hoc Tukey test