Fig. 3

LRRK2 interacts with CELSR1 and PRICKLE1, two key components of WNT/PCP signaling. a-d Western blotting analysis of co-IP of transiently overexpressed LRRK2 with WNT/PCP signaling components in HEK293T. LRRK2 physically interacts with CELSR1 (a) and PRICKLE1 (b), but neither with VANGL2 (c) nor ROR2 (d). e-i IF of proteins overexpressed in HEK293T. Under normal conditions, LRRK2 is evenly distributed in the cytoplasm (i). CELSR1 alone is usually polarized in cells (f), whereas PRICKLE1 tends to form puncta (h). e Once co-expressed, LRRK2 partially co-localizes with CELSR1 in the cytoplasmic membrane and in cell-cell contacts (arrowheads). LRRK2 robustly changes its localization when it is co-expressed with PRICKLE1, and together they form puncta structures in the cytoplasm (g). Arrowheads show co-localizations, whereas arrows point out that there is no leakage of the fluorescent signal in PRICKLE1-positive and LRRK2-negative cells. Nuclear staining by Dapi is in blue. N ≥ 3. Scale bars show 20 μm. j-l Physical interaction between LRRK2 and CELSR1 (j) or PRICKLE1 (k-l) is not modified in the most common LRRK2 mutations. l Analysis of immunoblot band intensity shows the relative binding of LRRK2 WT and mutants to PRICKLE1. No difference between LRRK2 WT and mutants was detected. Signal was adjusted to the background and normalized to the corresponding input (N = 3, SD)