Figure 5. ES cell proliferation rescue by wild-type CITED2 and CITED2-T166N in the absence of leukemia inhibitory factor (LIF).

E14/T ES cells transfected by electroporation with the indicated plasmids were selected in puromycin. Surviving cells were plated in quadruplicate into 12-well plates in medium containing puromycin, but in the absence of LIF. The cells were fixed at the indicated time points using 10% formalin. The relative number of ES cells was determined by staining with 0.1% crystal violet, extracting cell-associated dye using 10% acetic acid, and measuring absorbance at A590. Results are presented as mean±SEM. Results represent a single experiment. Similar results were obtained from three independent experiments.