Protocol to quantify LRRK2-mediated Rab10 phosphorylation using high-content imaging.

Authors

Beatrice Masotti, Paolo Lorenzon, Giuseppina Covello, Elisa Greggio

Published in

STAR protocols. Volume 7. Issue 3. Pages 104819. Sep 04, 2026. Epub Sep 04, 2026.

Abstract

Rab10 phosphorylation at Thr73 (pRab10) is a well-established readout of the kinase LRRK2, a protein whose mutations are associated with Parkinson's disease. Here, we present a protocol for high-content quantification of endogenous pRab10-positive vesicles in primary astrocyte-enriched cultures using the Operetta CLS system. We describe steps for cell culture, immunofluorescence, image acquisition, and quantitative analysis using Harmony software. This protocol is applicable to other LRRK2-expressing cells and offers a scalable platform for quantitative studies of LRRK2 physiological and pathological activity.

PMID:
42696457
Bibliographic data and abstract were imported from PubMed on 05 Sep 2026.

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