Fig. 4.

ACR16 and PRE-084 increased synapsin1 protein expression and activated both MAPK and PI3K signaling pathways, in which ACR16 maintained this activation over a prolonged period. Rat hippocampal primary cell cultures were treated with 70 nM ACR16 or 50 nM PRE-084 at 12 DIV, and the phosphorylation status of AKT and ERK 1/2 was quantified using Western blot (a and b), in addition to protein expression of synapsin1 (c). An extended time-course, from 15 min to 72 h, was used to study short and longer cellular events. Data were normalized to control conditions and expressed as the percentage change of pAKT (a) or pERK 1/2 (b) normalized to pan AKT or ERK1/2 protein respectively, as well the percentage change of protein expression of synapsin1 normalized to pan AKT (c). 2-way ANOVA followed by Sidak’s multiple comparisons test was used for data analysis, comparing the percent change over time versus untreated control basal levels at time point 0, as well as between treatments. Significant differences compared with untreated controls are displayed as *(p < 0.05), **(p < 0.01), and ***(p < 0.001), respectively. Significant differences between ACR16 and PRE-084 treatments are shown as #(p < 0.05) and ##(p < 0.01). Representative Western blots are different time points are presented in d and e