Fig. 2

Selection and primary tests of mouse ESCs bearing the developed Oct4 tracing system (O4S ESCs). a PCR-genotyping of O4S ESC line showing that constructs were inserted in the correct genome locations (left panel); normal chromosome counts of O4S ESCs (right panel); results of the sequencing of targeted loci, showing the correct position of the inserted cassette (bottom panels). b Verification of pluripotency status of the generated O4S ESCs. Comparison of the pluripotency marker expression in O4S vs. wild-type (WT) ESCs by flow cytometry (left and top panels) and qRT-PCR (chart); results are expressed as mean ± SD. Teratoma assay showing the ability of O4S ESCs to differentiate into three germ layers (bottom panels). c The ability O4S ESCs to activate the tdTomato expression upon the administration of both 4-OHT and Dox. Representative flow cytometry showing distinct ability to activate tdTomato expression in O4S vs. O4S-tTR ESCs which are different only the tTR-encoding sequence in the targeted Oct4 locus