Fig. 5.

Dephosphorylation at residues S908, S910, S935, S955, S973, or S976 is not crucial for LRRK2 kinase inhibitor-induced destabilization. SH-SY5Y cells overexpressing LRRK2 with (a, c) 6xSA (S908A/S910A/S935A/S955A/S973A/S976A) or (b, d) 6xSE (S908E/S910E/S935E/S955E/S973E/S976E) were treated (a, b) for different periods of time with PF-06447475 (150 nM), MLi-2 (10 nM), or DMSO as control or (c, d) between 6 and 8 h with MLi-2 (10 nM), IC261 (300 μM), or DMSO as control. Cell lysates were analyzed with immunoblotting using FlagM2 antibody for LRRK2 detection or anti-vinculin for equal loading. Representative blots are shown. Graphs show the quantification of blots representing the ratio of total LRRK2 over housekeeping protein signal. Error bars indicate S.E.M. with N ≥ 3. Statistical significance was tested using a 2-way ANOVA test with Bonferroni post-tests or column statistics with Bonferroni correction. Triple asterisks indicate p < 0.001, asterisk indicates p < 0.05