Author response image 1. Quantitative analysis of VPS35 knockdown efficiency.
(A) HEK 293 cells were transfected with lentiviral vector expressing siRNA along with EGFP followed by immunoblotting with antibodies to VPS35 and β-actin after 72 h. Right panel shows quantification of immunoblots. Relative amount of VPS35 was determined with NIH ImageJ (N = 3 experiments). (B) Hippocampal neurons were transfected with lentiviral vector expressing siRNA along with GFP at DIV12, fixed on DIV18 and immunostained with antibodies to VPS35. Right panels are quantification of VPS35 signals in the outlined area of neurons (mean ± SEM, n = 10 neurons from three independent experiments). Scale bar, 5 μm.