Fig. 2.
Protein levels of truncated forms of LRRK2 are not reduced upon LRRK2 kinase inhibitor treatment. We generated SH-SY5Y cells overexpressing 3Flag-truncated forms of LRRK2 (a): LRRK2823-2527 or PLRCKW, LRRK2702-2527, or APLRCKW. Cell lines were either not treated (b) or treated for different periods of time with PF-06447475 (150 nM), MLi-2 (10 nM), or DMSO as control (c). Cell lysates were analyzed with immunoblotting using FlagM2 antibody for LRRK2 detection, anti-LRRK2 P-S935, anti-LRRK2 P-S1292, 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 or the ratio of phosphorylation at S935 over total LRRK2 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, double asterisks indicate p < 0.01