Additional characterization of the LRRK2-TRIM1 interaction. (a–c) Live-cell confocal microscopy of GFP-LRRK2 and mCherry-tubulin or mCherry-TRIM1 transiently transfected into A549 cells (scale bar = 10 μM; a), SK-N-SH cells (scale bar = 5 μM; b) or HEK-293T cells (scale bar = 5 μM; c). From top to bottom, each set shows mCherry-tubulin or mCherry-TRIM1, GFP-LRRK2, merged image. In all lines examined, in the presence of mCherry-tubulin, GFP-LRRK2 is diffusely cytoplasmic, but microtubule-localized in the presence of mCherry-TRIM1. (d) Alignment of TRIM18 with TRIM1. Domains labeled above alignment. Red line designates region required for TRIM1 interaction with LRRK2. Double red line designates region of least homology in TRIM1 and TRIM18 dual B-box domain. Dual-AAA motifs below the sequence designate the mutated amino acids used to make cytoplasmic TRIM1 C variant. (e) Immunoprecipitation of GFP-LRRK2, which fails to coimmunoprecipitate with HA-TRIM9 in HEK-293T cells. Source data are available for this figure: SourceData FS1.