(A) RPE cells were ciliated for 24 hr and stained with rabbit anti-RILPL1 antibodies (red) and co-stained with mouse anti-Arl13b or rabbit anti-Rab10 antibodies (green). (B) Rat primary astrocytes were ciliated and stained with anti-RILPL1 and anti-Arl13B antibodies as indicated. (C–E) HEK293T cells were infected with lentiviral shRNA for RILPL1 and selected by puromycin. LRRK2-R1441G was co-transfected with either RILPL1-WT. Also, RILPL1-C-terminus GFP or RILPL1-N-terminus GFP (green) were co-transfected with LRRK2 in wildtype cells. After 24 hr, cells were stained with anti-phospho-Rab10 antibody (red). (D) Quantitation of cells with peri-centriolar clustering of pRab10 in RILPL1 knock down HEK293T cells, with or without rescue by (full length) RILPL1-WT. (E) Quantitation of peri-centriolar clustering of pRab10 in cells transfected with full length, RILPL1-N terminus, RILPL1-C terminus. (F) A549 WT and A549-RILPL1 knock out cells were transfected with myc-LRRK2-R1441G and 24 hr later, stained with anti-pRab10 (red) and DAPI (blue). (G) Quantitation of total pRab10 in peri-centriolar clusters. (H) A549 WT and A549-RILPL1 knock out cells were lysed and analyzed by immunoblotting with the indicated antibodies. Error bars represent SEM from three experiments with >50 cells per condition in each experiment. *, p<0.05; **, p<0.05; ***, p<0.001. Scale bar, 10 µm.