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. 2022 Jul 8;12(5):1423–1447. doi: 10.3233/JPD-213128

Fig. 6.

Fig. 6

Immunocytochemistry and western blotting of control and G2019S LRRK2 LCLs using various antibodies. A) Example of control (ctrl) WT and G2019S LRRK2 LCLs stained with anti-LRRK2 antibody (N241A/34) and DAPI. B) Example of LCLs stained with knockout-validated RAB10 (SAB5300028) antibody (green), centrosomal marker (pericentrin, red) and DAPI. C) Example of LCLs stained with sheep anti-pT73-RAB10 antibody (green), centrosomal marker (pericentrin, red) and DAPI. The sheep antibody was either preabsorbed with a 10-fold molar excess of dephospho-peptide or of phospho-peptide as indicated. Identical results were obtained when employing the rabbit monoclonal anti-pT73-RAB10 antibody (Abcam, ab241060) [20]. D) Cells were incubated in the absence or presence of MLi-2 (100 nM, 2 h), and extracts subjected to western blotting using the indicated antibodies. E) Example of cells in absence or presence of MLi-2 (10 nM, 2 h), and stained with pS1292-LRRK2 antibody (Triton-X100/PBS-containing buffer conditions) and DAPI. F) As in (E), but cells stained with pS1292-LRRK2 antibody (PLA buffer conditions) and DAPI. Scale bars, 10 μm. Representative western and images are from two independent experiments, with comparable results obtained in both cases.