Figure 4.
E193K variant affects LRRK2 supra-molecular organization. (A) Protein lysate prepared from primary fibroblast obtained from healthy individuals or E193K carrier was separated by size exclusion chromatography (SEC). LRRK2 elution profile was revealed by dot-blot using anti-LRRK2 antibody. Theoretical molecular weight are: V0 at fraction 8.5, 669 kDa at fraction 12, 449 kDa at fraction 13. (B) The graph reports the intensity of each dot (fraction) normalized by the integrated intensities. Data are shown as mean ± SE, n = 4; *p < 0.05 vs. WT. Column void volume is 7.5 ml. (C) Primary fibroblasts obtained from healthy individuals or an E193K carrier were assayed by filter retardation assay to isolate high molecular weight (HMW) form of LRRK2 on acetate cellulose membrane and total LRRK2 on nitrocellulose membrane. (D) The graph reports LRRK2 HMW amount expressed as fold over total LRRK2. Data are shown as mean ± SE, n = 8; **p < 0.01 vs. WT.