EWS/WT1-KTS and EWS/WT1 + KTS enhances clonogenicity in p53
MEFs in reduced serum. (A) Western blot analysis of lysates derived from wild-type, p53+/-, and p53-/- background MEFs which were infected with doxycyline repressible eGFP, EWS/WT1-KTS or EWS/WT1 + KTS lentiviral particles, showing expression of EWS/WT1 in the absence of doxycycline. (B) Representative images from one experiment showing MEF colonies of the indicated genotype expressing eGFP, EWS/WT1-KTS or EWS/WT1 + KTS, plated at 1× 105 cells per plate and cultured in 1% or 2% fetal calf serum. After 14 days cells were fixed with glutaraldehyde and stained with crystal violet. (C) Mean number of colonies (from three independent experiments) greater than 5 mm of cells cultured in 1% (left panel) or 2% (right panel) serum on day 14. Error bars show ± SEM from three independent experiments. # denotes a p value of <0.05 as determined by Student’s t-test comparing eGFP to EWS/WT1-KTS and eGFP to EWS/WT1 + KTS.