A. Protein lysates from the brain (upper panels), kidney (middle panels) or lung (lower panels) of WT or E2385R knockin mice. Each lane is a separate animal. In each tissue, cyclophilin B was used as a loading control.
B. Quantification of steady state of mouse Lrrk2 levels relative to cyclophilin B in each genotype and tissue. In kidney tissue, the double bands detected as Lrrk2 were each quantified as one band. Error bars indicate SEM, n=4 animals per genotype.
C. Protein lysates from the brain (left panel) or lung (right panel) of WT or E2385R knockin mice. Each lane is a separate animal. Cyclophilin B was used as a loading control.
D. Quantification of phospho-serine 935 and phospho-serine 1292 relative to total Lrrk2 protein in each genotype and tissue. Two-way ANOVA indicated that there was a statistically significant difference in pS935 and pS1292 phosphorylation between tissues but not between genotypes (pS935: F tissue (1, 12) = 52.51, p<0.0001; F genotype (1, 12) = 2.645, p=0.1298. pS1292: F tissue (1, 12) = 53.29, p<0.0001; F genotype (1, 12) = 0.2364, p=0.6356; n = 4 animals per genotype).
E. Protein lysates from the brain (left panel) or lung (right panel) of WT or E2385R knockin mice. Each lane is a separate animal. Cyclophilin B was used as a loading control.
F. Quantification of phospho-T72 Rab8A and phospho-T73 Rab10 relative to total Rab protein in each genotype and tissue. Two-way ANOVA indicated that there was a statistically significant difference in Rab8a phosphorylation between tissues but not between genotypes (F tissue (1, 12) = 18.07, p=0.001; F genotype (1, 12) = 0.2564, p=0.6218; n=4 animals per genotype). Similarly, twoway ANOVA indicated that there was a statistically significant difference in Rab10 phosphorylation between tissues but not between genotypes (F tissue (1, 12) = 45.55, p<0.0001; F genotype (1, 12) = 0.239, p=0.6337; n = 4 animals per genotype).