Proteomics analysis of differentially expressed proteins by p53 status and cholesterol concentration of culture medium in on-top 3D culture using MDA-MB-231

Tumor cells had p53 knocked down by CRISPR/Cas9. In addition, Doxycycline-induced p53WT or p53R280K was incorporated into the p53KO cells by the Tet-on system. Furthermore, we created a high cholesterol condition in that Cholesterol was added to the culture medium while p53R280K was being expressed, and a low cholesterol condition in which Cholesterol was reduced by reducing serum. The cells were cultured for 7 days in on-top 3D with respect to these four conditions. Whole cell lysates were processed for non-targeted proteomic analysis.