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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: Parkinson’s disease-associated LRRK2-G2019S mutant acts through regulation of SERCA activity to control ER stress in astrocytes

Fig. 3

LRRK2-GS interacts with SERCA. a HEK293T cells were co-transfected with siRNA targeting the 3′-UTR region of LRRK2 and 3xMyc-tagged wild-type LRRK2 or G2019S-mutated LRRK2. After 48 h, cells were stimulated with α-synuclein (α-Syn) for 48 h and immunoprecipitated with antibody against Myc. LRRK2-interacting proteins were identified by MS analysis (Additional file 1: Table S1). This analysis revealed that LRRK2-GS–interacting proteins were enriched for Gene Ontology (GO) Biological Process terms associated with the ER stress response compared with LRRK2-WT–interacting proteins (> 1.5 fold, p-value < 0.05). Heatmap illustrating enriched LRRK2-GS–interacting proteins from three independent samples. The color bar represents fold change. b, c Astrocytes isolated from non-Tg and LRRK2-GS mice were treated with α-Syn for 48 h. Protein interactions were measured by immunoprecipitation with an antibody against LRRK2 or SERCA (b), and LRRK2–SERCA interactions were detected using the PLA method (c). PLA signals (in gray) were quantified, and displayed graphically as the average number of spots. Data are presented as means ± SD of three independent experiments (*p < 0.05). d HEK293T cells co-transfected with the indicated constructs were immunoprecipitated with an anti-Flag antibody, followed by immunoblot analysis with the indicated antibodies. Schematic diagram showing the structure of Flag-LRRK2

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