Figure 6 | Scientific Reports

Figure 6

From: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

Figure 6The alt text for this image may have been generated using AI.

In HEK293 cells exogenous LRRK2 phosphorylates Rab12 at Ser106 in a kinase activity-dependent way. (A) Odyssey CLx scan Western Blot image showing overexpressed total LRRK2 and HA-tagged Rab10 immunoreactivity (red panels), LRRK2-pSer935 and Rab12-pSer106 immunoreactivity (green panels) as well as overlay in cells overexpressing various LRRK2 exonic variants with either HA-tagged wild type Rab12 or HA-tagged phosphodeficient Rab12-S106A. Top panel, Rab12 and Rab12-pSer106. Bottom panel, LRRK2 and LRRK2-pSer935. Full-length blots are presented in Supplementary Figure 3. (B) Quantification of normalized Rab12-pSer106/total Rab12 ratio (n = 3 experiments). (C) Quantification of normalized LRRK2-pSer935/total LRRK2 ratio (n = 3 experiments). (D) Odyssey CLx scan Western Blot image example showing overexpressed total LRRK2 and HA-tagged Rab12 immunoreactivity (red panels), LRRK2-pSer935 and Rab12-pSer106 immunoreactivity (green panels) as well as overlay in cells overexpressing various LRRK2 exonic variants with HA-tagged wild type Rab12 and treated with either the LRRK2 inhibitor Lu AF58786 or DMSO. Top panel, Rab12 and Rab12-pSer106. Bottom panel, LRRK2 and LRRK2-pSer935. Full-length blots are presented in Supplementary Figure 3. (E) Quantification of normalized Rab12-pSer106/total Rab12 ratio (n = 3 experiments). (F) Quantification of normalized LRRK2-pSer935/total LRRK2 ratio (n = 3 experiments). Data was analyzed by two-way ANOVA with Holm-Sidak’s multiple comparisons test. Data presented as means ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 vs. controls (Rab12 or DMSO).

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