Retroviral Transduction of Tsc2+/− Eμ-Myc Lymphomas

Tsc2^+/−Eμ-Myc lymphomas were maintained in B-cell media (45 % DMEM, 45 % IMDM, 55 μM β-mercaptoethanol and 10 % fetal bovine serum) on γ-irradiated Arf^−/− MEF feeder layers. Retroviral packaging was performed using ecotropic Phoenix cells according to established protocols (http://web.stanford.edu/group/nolan/_OldWebsite/protocols/pro_helper_dep.html). Tsc2^+/−Eμ-Myc lymphomas were infected with MLS retrovirus expressing shFLuc.1309 as neutral control [21 (link)] or shMcl-1.1334 [22 (link)]. The amount of GFP⁺ cells was determined 12 h after transduction (t = 0) and again 15 h later by flow cytometry using a Guava EasyCyte HT FACScan instrument and Guava ExpressPro software (Millipore).

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Nguyen M., Cencic R., Ertel F., Bernier C., Pelletier J., Roulston A., Silvius J.R, & Shore G.C. (2015). Obatoclax is a direct and potent antagonist of membrane-restricted Mcl-1 and is synthetic lethal with treatment that induces Bim. BMC Cancer, 15, 568.

Publication 2015

Guava EasyCyte HT FACScan Guava ExpressPro software

B cell Feeder layers Fetal bovine serum Flow cytometry Guava Lymphomas Mercaptoethanol Retrovirus Tsc2

Corresponding Organization :

Other organizations : McGill University

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Variable analysis

independent variables

Retroviral expression of shFLuc.1309 or shMcl-1.1334

dependent variables

Amount of GFP+ cells at 12 hours (t = 0) and 15 hours after transduction

control variables

Tsc2+/- Eμ-Myc lymphomas maintained in B-cell media on γ-irradiated Arf-/- MEF feeder layers
Retroviral packaging performed using ecotropic Phoenix cells

positive controls

MLS retrovirus expressing shFLuc.1309 as neutral control

negative controls

No negative controls explicitly mentioned

Annotations

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