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. 2012 Oct 12;22(2):328–344. doi: 10.1093/hmg/dds431

Figure 2.

Figure 2.

Effect of LRRK2-IN1 and TTT-3002 on S935 phosphorylation of LRRK2 in human cell lines. LRRK2-IN1 and TTT-3002 inhibited S935 phosphorylation of LRRK2 in human THP-1 macrophages (A), human lymphoblastoid cell line derived from a PD patient carrying the G2019S mutation (B) and human SH-SY5Y neuroblastoma cell line transfected with G2019S-LRRK2 plasmid (C). The inhibitory effects of LRRK2-IN1 and TTT-3002 on S935 phosphorylation of LRRK2 were attenuated in cells transfected with G2019S/A2016T plasmid (D). All cell lines were treated with varying concentrations of LRRK2-IN1 and TTT-3002 for 1.5 h. Cell lysates were subjected to immunoblotting for detection of S935 phosphorylation (pS935 LRRK2) using a rabbit mAb UDD2 against phospho-serine 935 LRRK2, and for detection of total LRRK2 using a mouse mAb against LRRK2 amino acids 970–2527. The position of the 250 kDa molecular weight marker was indicated on each blot. The bar graphs represent percent pS935 LRRK2 normalized to total LRRK2 in untreated and inhibitor-treated samples, using band intensities determined from scanned blot images with the ImageJ software. The results were obtained from three independent experiments and values are expressed as mean ± SEM.