Figure 2
Validation after Cre-excision in mESCs and hematopoietic cells. (A) Schematic representation of Cre-recombinase mediated removal the floxed transcriptional stop cassette, leading to expression of the putative oncogene (in this case Jarid2) and the eGFP /luciferase reporter (eGFP/Luc; fusion protein). (B) Firefly-Luciferase activity in parental (P) and Cre-excised (CREX) RMCE-DV3 mESCs. For each putative oncogene, we Cre-excised two independent parental clones, and for each clone we measured luciferase activity in four parental and 10 Cre-excised mESCs. (C–F) qRT-PCR for four putative oncogenes, namely Jarid2, Runx2, dnETV6 and MN1 in their respective conditional KI mice. In addition, the presence of the floxed stop cassette and of eGFP/Luc was analyzed (C) qRT-PCR analysis in splenocytes of 11w old Cre + ve R26-Jarid2tg/tg; VaviCretg/+ (JV+) and Cre-ve (JV−) mice. The average and standard deviation are shown for three JV+ and three JV− spleens. (D) qRT-PCR analysis in fetal livers (FLs) of 15.5 dpc old Cre + ve R26-Runx2tg/tg; VaviCretg/+ (RV+) and Cre-ve (RV−) embryos. The average and standard deviation are shown for two RV+ and two RV− FLs. (E) qRT-PCR analysis in splenocytes of 8w old Cre + ve R26-dnETV6tg/tg; VaviCretg/+ (dEV+) and Cre-ve (dEV−) mice. The average and standard deviation are shown for two dEV+ and two dEV− spleens. (F) qRT-PCR analysis in splenocytes of 11w old Cre + ve R26-MN1tg/tg; VaviCretg/+ (MV+) and Cre-ve (MV−) mice. The average and standard deviation are shown for four JV+ and four JV− spleens.
