Figure S2.
Latex bead assay for CASM and validation of LRRK2 inhibitor. (A) Representative timelapse widefield microscopy images of MCF10A cells expressing GFP-LC3A and stained for LAMP1 (red), following engulfment of 3 μm latex beads and treatment with LLOMe (250 μM, 30 min) ± BafA1 (100 nM). Cropped images show LAMP1-positive latex-bead containing phagosomes. Scale bar, 10 μm. (B) Quantification of GFP-LC3A recruitment to LAMP1 positive phagosomes. Data represent mean ± SEM from three independent experiments. ****, P < 0.0001, unpaired t test. (C) Western blot analysis of LRRK2 substrate, Rab10, in ATG13 KO MCF10A cells treated with LLOMe (250 μM, for 20 min or 4 h) ± LRRK2 inhibitor MLi2 (1 μM, 1.5 h pretreatment). Samples probed for Rab10 and phospho T73 Rab10. Source data are available for this figure: SourceData FS2.

Latex bead assay for CASM and validation of LRRK2 inhibitor. (A) Representative timelapse widefield microscopy images of MCF10A cells expressing GFP-LC3A and stained for LAMP1 (red), following engulfment of 3 μm latex beads and treatment with LLOMe (250 μM, 30 min) ± BafA1 (100 nM). Cropped images show LAMP1-positive latex-bead containing phagosomes. Scale bar, 10 μm. (B) Quantification of GFP-LC3A recruitment to LAMP1 positive phagosomes. Data represent mean ± SEM from three independent experiments. ****, P < 0.0001, unpaired t test. (C) Western blot analysis of LRRK2 substrate, Rab10, in ATG13 KO MCF10A cells treated with LLOMe (250 μM, for 20 min or 4 h) ± LRRK2 inhibitor MLi2 (1 μM, 1.5 h pretreatment). Samples probed for Rab10 and phospho T73 Rab10. Source data are available for this figure: SourceData FS2.

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