TSC2^+/-Eμ-Myc and Eμ-Myc lymphomas are seeded in 96-well plates at 10⁶ cells/mL in the presence of increasing concentrations of doxorubicin (Dxr) (ranging from 3.9 nM to 250 nM) and 4E1RCat (ranging from 78.13 nM to 10 000 nM) at a constant ratio of either 20:1 or 40:1. Twenty four hours later, a MTS assay is performed. To this end, Cell Proliferation Assay is added to the plates and the plates further incubated for up to 3 h, followed by measuring the OD₄₉₀. Values obtained are standardized against DMSO controls^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
One million secondary Pten^+/-Eμ-Myc, Tsc2^+/-Eμ-Myc, or Eμ-Myc lymphoma cells are injected into the tail vein of 6-8 week old female C57BL/6 mice. When tumors are palpable, mice are treated with rapamycin (4 mg/kg daily for 5 d), 4E1RCat (15 mg/kg daily for 5 d), or doxorubicin (once at 10 mg/kg). Compounds are administered via intraperitoneal (i.p.) injection in 5.2% PEG 400/ 5.2% Tween 80. For combination studies, rapamycin or 4E1RCat are injected i.p. daily for five consecutive days, with doxorubicin being administered once on day two. Animals are palpated daily to monitor for the onset of tumors. Tumor-free survival is defined as the time between disappearance and reappearance of tumors. Data is analyzed using the log-rank test for statistical significance presented in Kaplan-Meier format^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Cencic R, et al. Reversing chemoresistance by small molecule inhibition of the translation initiation complex eIF4F. Proc Natl Acad Sci U S A. 2011 Jan 18;108(3):1046-51.  [Content Brief]

  [2]. Shuda M, et al. CDK1 substitutes for mTOR kinase to activate mitotic cap-dependent protein translation. Proc Natl Acad Sci U S A. 2015 May 12;112(19):5875-82.  [Content Brief]