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. 2017 Dec 4;7:16827. doi: 10.1038/s41598-017-17119-1

Figure 5.

Figure 5

Generation and evaluation of Dicer −/−;Pten −/−;Trp53 −/− TKO cells. (A) OvidT 497 Dicer −/−;Pten −/− (DKO) cells were transfected with PX459 encoding Trp53 gRNA. Clone 4 contained no Trp53 mutation; clone 13 (TKO) contained bi-allelic Trp53 exon 5 mutations. Expression of PTEN and p53 was assessed by immunoblot (left). F3 = ID8 Trp53 −/−. Sensitivity to Nutlin-3 was assessed by MTT assay (right). (B) Homologous recombination was assessed in DKO 4 and TKO 13 cells as previously. (C) Sensitivity of DKO 4 and TKO 13 cells to cisplatin. Each dot represents one triplicate experiment. Bars represent median. *p < 0.01. (D) Cells (5 × 106) were injected intraperitoneally into female C57Bl/6 mice in groups of six. Mice were killed when they reached humane endpoints. Excised tumours were fixed in formalin and stained for WT1 and PAX8. Each TKO 13 section comes from a separate mouse. Positive controls (+ve) are ID8 tumour (WT1) and normal mouse fallopian tube (PAX8), both from14. Bars represent 50 µm.