Isolation and Characterization of Mouse Keratinocytes and MEFs

Primary keratinocytes and early passage MEFs from Mdm2^SNP309G/G and Mdm2^SNP309T/T mice were generated as previously described (21 (link), 55 (link)). Early passage MEFs were transfected with 7 µg of pCMV-GFP vector containing E2F4 cDNA, E2F6 cDNA, SP1 cDNA, or empty vector using Lipofectamine 3000 reagent (Sigma, L3000015). Forty-eight hours later, GFP positive cells were sorted by FACS and RNA was extracted for RT-PCR analysis.

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Ortiz G.J., Li Y., Post S.M., Pant V., Xiong S., Larsson C.A., El-Naggar A.K., Johnson D.G, & Lozano G. (2017). Contrasting effects of an Mdm2 functional polymorphism on tumor phenotypes. Oncogene, 37(3), 332-340.

Publication 2017

Lipofectamine 3000 reagent

Cdna Cells Keratinocytes Lipofectamine Mice Rt pcr Vector

Corresponding Organization :

Other organizations : The University of Texas MD Anderson Cancer Center, University of Houston

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Variable analysis

independent variables

Mdm2 genotype (Mdm2SNP309G/G and Mdm2SNP309T/T)
Transfection with pCMV-GFP vector containing E2F4 cDNA, E2F6 cDNA, SP1 cDNA, or empty vector

dependent variables

Gene expression (measured by RT-PCR analysis)

control variables

Primary keratinocytes and early passage MEFs
Lipofectamine 3000 reagent for transfection
Sorting of GFP positive cells by FACS

controls

Negative control: Empty vector transfection

Annotations

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