Effect of MDM2 overexpression on response to etoposide treatment
We aimed to study the effect of non-mutational p53 inactivation on the response of hematopoietic progenitor cells to genotoxic treatment. In order to study this in vitro, we generated HSPC lines by enforced ER-Hoxb8 expression in bone marrow cells derived from transgenic Trp53-fl-R245W-GFP mice. In the absence of Cre-mediated recombination, these cells are wild-type for Trp53. We additionally transduced these cells lentivirally with a construct overexpressing murine Mdm2 or a control vector. This leads to non-mutational inactivation of functional p53. Thereby, we aimed to compare the transcriptional response to genotoxic treatment in Trp53 wild-type cells with or without non-mutational p53 inactivation through Mdm2 overexpression Overall design: Hoxb8 cells, immunophenotypically resembling murine granulocyte-macrophage progenitor cells, were transduced with either an Mdm2 overexpressing or a control vector. These cells were treated with 10 uM etoposide for 3 hours, after which RNA was isolated and subjected to bulk RNA sequencing.