Pharmacological Inhibition of LRRK2, Rab7, and PIKfyve

All drugs used in this study were dissolved in DMSO, diluted into culture medium and the medium sterile filtered prior to use. The LRRK2 kinase inhibitors LRRK2-In1 and GSK2578215A were from Merck and R&D Systems, respectively. The Rab7 GTPase inhibitor, CID 1067700 (Agola et al., 2012 (link)) was from EMD Millipore. The NAADP antagonist trans-Ned-19 was synthesised as described previously (Naylor et al., 2009). Ned-K is an analogue of Ned-19 in which the fluoride has been replaced with a cyano group. Its synthesis will be described elsewhere. Both Ned-19 and Ned-K were kind gifts from A. Ganesan (School of Pharmacy, University of East Anglia, UK), Raj Gossain (School of Chemistry, University of Southampton, UK) and Sean M. Davidson (Hatter Institute, UCL, UK). The PIKfyve inhibitor, YM-201636 was from Cambridge Bioscience. BAPTA-AM and EGTA-AM were from Sigma.

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Hockey L.N., Kilpatrick B.S., Eden E.R., Lin-Moshier Y., Brailoiu G.C., Brailoiu E., Futter C.E., Schapira A.H., Marchant J.S, & Patel S. (2015). Dysregulation of lysosomal morphology by pathogenic LRRK2 is corrected by TPC2 inhibition. Journal of Cell Science, 128(2), 232-238.

Publication 2015

LRRK2-In1 BAPTA-AM EGTA-AM

Bapta Cid 1067700 Dmso Drugs Egta Fluoride Gifts Gsk2578215a Gtpase Inhibitors Kinase Lrrk2 in1 Naadp Ned 19 Sterile Synthesis Ym 201636

Corresponding Organization :

Other organizations : University College London, University of Minnesota Medical Center, Thomas Jefferson University, Temple University

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Variable analysis

independent variables

LRRK2 kinase inhibitors LRRK2-In1 and GSK2578215A
Rab7 GTPase inhibitor, CID 1067700
NAADP antagonist trans-Ned-19
Ned-K (analogue of Ned-19)
PIKfyve inhibitor, YM-201636

dependent variables

Unspecified

control variables

All drugs were dissolved in DMSO, diluted into culture medium and the medium sterile filtered prior to use
BAPTA-AM and EGTA-AM were used

positive controls

Unspecified

negative controls

Unspecified

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