Protocol detail

Molecular Tools for Cell Line Analysis

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Cell lines HEK293T (ATCC), Hs578T (ATCC), and MDA-MB-231LN (Caliper LifeScience) were grown based on the manufacturer's recommendations. shRNAs and siRNAs against NEDD9, non-targeting control (ThermoFisher Scientific), and NEDD9-cDNA were prepared as previously described (7 (link)). mRFP-Rab5, and -Rab7 were purchased from Addgene (plasmid #14437, #14436) (60 (link)). Plasmids mCherry-MMP14 and PAmCherry-MMP14 were a gift from Dr.Chavrier (Institute Curie, France) and Dr.Machesky (The Beatson Institute for Cancer Research, Scotland) (24 (link)). Human TIMP2-cDNA (ThermoFisher Scientific) was sub-cloned into pcDNA3.1-3x-Flag plasmid (gift from Dr.Frisch, West Virginia University, USA).

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Loskutov Y.V., Kozyulina P.Y., Kozyreva V.K., Ice R.J., Jones B.C., Roston T.J., Smolkin M.B., Ivanov A.V., Wysolmerski R.B, & Pugacheva E.N. (2014). NEDD9/Arf6-Dependent Endocytic Trafficking of Matrix Metalloproteinase 14: A Novel Mechanism for Blocking Mesenchymal Cell Invasion and Metastasis of Breast Cancer. Oncogene, 34(28), 3662-3675.

Publication 2014

HEK293T Hs578T MDA-MB-231LN mRFP-Rab5, Human TIMP2-cDNA

Cancer Cdna Cell lines Human Mmp14 Plasmid Shrnas Sirnas Timp2

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Protocol cited in 1 other protocol

Variable analysis

independent variables

ShRNAs and siRNAs against NEDD9
NEDD9-cDNA
MRFP-Rab5
MRFP-Rab7
MCherry-MMP14
PAmCherry-MMP14
TIMP2-cDNA

dependent variables

Not explicitly mentioned

control variables

Non-targeting control shRNAs and siRNAs

controls

Positive control: Non-targeting control shRNAs and siRNAs
Negative control: Not specified

Annotations

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