Fig. 4. DDX11 knockout causes TP53 activation and increased sensitivity to RAD51 inhibition.

a RPE1-hTERT cells and RPE1-hTERT-TP53KO cells, both containing a doxycycline inducible Cas9 construct, were transfected with DDX11 guide RNA and indels were analyzed using Sanger sequencing. More detailed information on guide RNA design and validation of clones is provided in Supplementary Fig. 5. b Cells were transfected with indicated siRNA’s, lysed after two days and analyzed by western blot. Note that DDX11KO cells have elevated p53 levels. A representative of two independent experiments is shown. c In parallel with b, cells were transfected with siRNA and proliferation was monitored using IncuCyte software. UBB siRNA was used to control transfection efficiency. A representative of two independent experiments is shown, with three technical replicates. Note that sip53 specifically accelerates growth of wtTP53-DDX11KO cells. d RPE1-hTERT and RPE1-hTERT-DDX11KO cells were transfected with indicated siRNAs. After two days, mRNA levels were assessed with qRT-PCR in three technical replicates. e RPE1-hTERT cells were cultured in a 96-wells plate in the presence or absence of the RAD51 inhibitor BO-2 (10 µM). Growth was monitored using IncuCyte software. In total six replicates from two independent experiments are shown.