Fig. 5

Synapsin 1 levels are decreased in the presence of A30P-aSyn oligomers in an NFAT-dependent pathway. Mouse primary hippocampal neurons (A) or differentiated N2a cells (C) were treated with A30P-aSyn-O (10 µM) in the absence or in the presence of VIVIT (10 µM) for 18 h. Genes responsive to NFAT pathway modulation found in the panel (Syn1, Tubb4, Kalrn, Bcl-2A1A and Bim) were validated by real time PCR. CA-NFAT-ER N2a cells were treated with 150 of OHT (Tamoxifen) and were analyzed after 24 h by real-time-PCR for the same genes (E). Western blot analyses were made for the same cells and treatments to assess expression level of Syn1 protein (B, D, F; left). β-Actin was used as loading control. Graphical representation of band intensities of Syn1 normalized to β-actin (B, D, F; right). Mouse primary hippocampal neurons were treated with 10 µM of A30P-aSyn-O in the absence or presence of 2 µM of CsA or 10 µM of VIVIT for 24 h. Cells were immunostained with anti-MAP2 and anti-synapsin-1 antibodies (G). All experiments were performed at least three times and graphs are represented as mean and SD where *p > 0.05, **p > 0.01 and ***p > 0.001. Scale bar = 10 μm