Cell Culture Conditions for RUNX1 Models

HeLa cells were cultured in DMEM medium (Thermo Fisher, Waltham, MA) supplemented with 10% FBS (Thermo Fisher). 32D cells expressing wild type RUNX1 and GCSF receptor [17 (link)] and 32D cells carrying the RUNX1-MTG8 fusion protein (32D-RM8) [13 (link)], and derived cell lines were cultured in RPMI 1640 (Thermo Fisher) medium supplemented with 10% heat-inactivated (HI) fetal bovine serum (FBS) (Thermo Fisher) and 1 ng/ml murine IL-3 (Peprotech, Inc., Rocky Hill, NJ). Cells were counted daily with a Coulter Particle Counter (Beckman Coulter, Brea, CA) and diluted to a density of 2 × 10⁵ cells/ml daily.

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Rossetti S., Anauo M.J, & Sacchi N. (2017). MiR-221-regulated KIT level by wild type or leukemia mutant RUNX1: a determinant of single myeloblast fate decisions that – collectively – drives or hinders granulopoiesis. Oncotarget, 8(49), 85783-85793.

Publication 2017

DMEM medium FBS RPMI 1640 murine IL-3 Coulter Particle Counter

Cell lines Cells Fetal bovine serum Gcsf receptor Hela cells Murine Runx1 Runx1 protein

Corresponding Organization : Roswell Park Comprehensive Cancer Center

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Variable analysis

independent variables

RUNX1 expression (wild type vs. RUNX1-MTG8 fusion protein)

dependent variables

Cell growth/proliferation (measured by daily cell counting)

control variables

Cell culture media (DMEM for HeLa cells, RPMI 1640 for 32D cells)
Serum supplements (10% FBS, 1 ng/ml murine IL-3 for 32D cells)
Cell seeding density (2 × 10^5 cells/ml)

controls

Positive control: 32D cells expressing wild type RUNX1 and GCSF receptor
Negative control: 32D cells carrying the RUNX1-MTG8 fusion protein (32D-RM8)

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