Quantification of LRRK2 and Rab29 in HEK293FT cells

HEK293FT cells were seeded at 2.5 × 10⁴ cells per well in a Matrigel-coated 96-well plate and transfected with 3xFlag-HD-LRRK2 and either 2xmyc-Gus, 2xmyc-Rab29 or mutant 2xmyc-Rab29 Q67L plasmid. 24 hours after transfection cells were fixed and stained, then imaged and analyzed on a high throughput Cellomics VTI arrayScanner as previously described [21 (link)].

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Langston R.G., Rudenko I.N., Kumaran R., Hauser D.N., Kaganovich A., Ponce L.B., Mamais A., Ndukwe K., Dillman A.A., Al-Saif A.M., Beilina A, & Cookson M.R. (2018). Differences in stability, activity and mutation effects between human and mouse Leucine-Rich Repeat Kinase 2. Neurochemical research, 44(6), 1446-1459.

Publication 2018

VTI arrayScanner

Cells Lrrk2 Matrigel Plasmid Transfection

Corresponding Organization : National Institute on Aging

Other organizations : National Institutes of Health

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Variable analysis

independent variables

3xFlag-HD-LRRK2
2xmyc-Gus
2xmyc-Rab29
Mutant 2xmyc-Rab29 Q67L

dependent variables

Cell phenotype

control variables

Cell seeding density (2.5 × 10^4 cells per well)
Matrigel-coated 96-well plate
Cell culture duration (24 hours after transfection)
Imaging and analysis method (Cellomics VTI arrayScanner)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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