α-Syn uptake by Oli-neu cells is dependent on clathrin expression and enhanced by exposure to PUFAs.

<p>A. Naïve Oli-neu were transiently transfected either with scramble or two different shRNAs designed against clathrin heavy chain. 56 hours post DNA-transfection, cells were cultured in standard growth medium supplemented with purified human α-Syn (at 15 µg/ml) for 16 hours and processed for ICC with anti clathrin and anti α-Syn antibodies. Bar, 10 µm. B. Graph representing quantitation of the signal obtained by ICC, mean ± SD of n = 20–30 cells in each treatment. *, p<0.001 ANOVA. C. Naïve Oli-neu cells (sister cultures) were incubated in the presence of purified α-Syn (at 15 µg/ml) added to standard growth medium (with serum); serum-free DMEM supplemented with BSA only (at 50 µM); or BSA together with the indicated FA (at 250 µM) for 16 hours. Cells were then washed and processed for ICC for the detection of intracellular α-Syn using anti the α-Syn antibody, LB509. The signal within cells was quantified. Mean ±SD of n = 10–15 cells in each treatment. *, P = 0.0034 (ANOVA).