Fig 1: The increase of DAT levels is associated with the G2019S mutation but not sustained by ongoing LRRK2 kinase activity. (A) Western blot analysis of DAT levels in the striatum of 12-month-old G2019S KI, LRRK2 KO and KD mice in comparison with WT controls, and representative blots. Data are means ± SEM of n = 6 mice per group * p < 0.05 different from WT (one-way ANOVA followed by the Tukey test). (B) Kinetic analysis of [3H]-DA uptake in striatal synaptosomes of 12-month-old G2019S KI and WT mice acutely treated with MLi-2 (10 mg/Kg i.p.) or vehicle. Data are expressed as mean ± SEM of n = 4 mice per group. (C,D) Western blot analysis of pSer1292 LRRK2 and total LRRK2 levels in the striatum of G2019S KI mice administered with MLi-2 (10 mg/Kg, i.p.) or vehicle, and representative blots. Data are expressed as mean ± SEM of n = 5 mice per group. ** p < 0.01 different from Vehicle (Student t-test two-tailed, for unpaired data).
Fig 2: Dysfunctional DAT activity in G2019S KI mice appears at 9 months of age. Kinetic analysis of [3H]-DA uptake in synaptosomes were performed in the striata of 6-month-old (A), 9-month-old (B), 12-month-old (C) and 18-month-old (D) G2019S KI mice versus age-matched WT controls. Data are expressed as mean ± SEM of n = 4 mice per group. Statistical analysis was performed using the Student t-test, two-tailed for unpaired data. * p < 0.05, different from WT mice.
Fig 3: Western blot data for TH (a,b), DAT (c,d), and DRD1 (e,f) for proton irradiated and control rats. Full-length blots/gels are presented in Supplementary Materials Figure S1a–c. *p < 0.05 compared with controls. N = 5–6/group.
Fig 4: Analysis of the dopaminergic synapse signaling pathway. (a) Western blotting showing the expression of DRD2, SLC6A3, and COMT proteins. GADPH was used as a loading control. (b), (c), and (d) The light blue bars represent the expression of DRD2, SLC6A3, and COMT in group C. The dark blue bars represent the expression of DRD2, SLC6A3, and COMT in group M. (e) and (f) mRNA expression of DRD2 and SLC6A3 in rat penile samples. Values are the mean ± standard error of the mean (n = 10 animals per group). The t-test was used. Group M was compared with group C, * represents P < 0.05.DRD2, dopamine receptor D2; SLC6A3, solute carrier family 6 member 3; COMT, catechol-O-methyltransferase; GAPDH, glycerol 3-phosphate dehydrogenase; C, control; M, model; OD, optical density.
Fig 5: Dopamine receptor and transporter levels by western blots: (A) DRD1, (B) DRD2, (C) DAT protein expression in hippocampus, nucleus accumbens, and striatum after DLM or CO. Data are averaged normalized signal relative to actin. Target protein is labeled in green and actin in red. Sample size: n = 7–10/treatment/sex/region. DA, dopamine; DRD1, dopamine D1 receptor; DRD2, dopamine D2 receptor; DAT, dopamine transporter; HIPP, hippocampus; NA, nucleus accumbens; STR, striatum. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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