Figure 5.

PNR502 blocks IL-1β-mediated increases in p38/MAPK and ApoE expression. (A,B) Primary neuronal cells, treated with IL-1β (30 ng/ml) in vitro, showed increased expression of p38/MAPK (quantified with antibody to the active form of p38/MAPK), but this response was blocked by pretreatment with 10-nM PNR502. The results are summarized in (B), showing means ± SD from three independent experiments, each comprising triplicate wells (as in A) and combined for statistical analysis by 2-tailed heteroscedastic t-test. (C) ApoE is induced by IL-1β treatment of NT2 cells (16 h, 30 ng/ml), but blocked if they are concurrently exposed to 10-nM PNR502; results, summarized in (D), show means ± SD from three independent experiments, each performed in duplicate. (E) Aβ_1–42- and tau-containing aggregates were isolated by immuno-pulldown (IP) as described previously (Ayyadevara et al., 2016b). These two aggregate types, and total aggregates (without IP) were resuspended in buffer containing 1% (w/v) sarcosyl, pelleted by high-speed centrifugation, recovered and analyzed by proteomics as described (Ayyadevara et al., 2016b). ApoE was roughly quantified in hippocampal aggregates from AD or controls (AMC), based on spectral hits (mean ± SEM) for four samples per group.