Skip to main content
. 2013 Feb 28;9(2):e1003292. doi: 10.1371/journal.pgen.1003292

Figure 3. Establishment of iPSC clones upon genome-wide erasure of H3K27me3 at the onset of reprogramming.

Figure 3

A. Western blot analysis of EZH2 and H3K27me3 protein levels respectively at onset or 48 hr after reprogramming in Ezh2+/ΔSET and Ezh2ΔSET/ΔSET MEFs. Data are representative of two experiments. Quantification of protein levels at the indicated time points is shown in the right panel (controls in grey, mutants in purple). B. Strategy to induce reprogramming of tail tip fibroblasts (TTFs) lacking H3K27me3 at the onset of reprogramming. C. Western blot analysis of EZH2 and H3K27me3 protein levels in Cdkn2a−/− TTF carrying either one (+/ΔSET) or both (ΔSET/ΔSET) Ezh2 mutant alleles. As comparison, representative Ezh2-proficient (+/+) and -deficient (ΔSET/ΔSET) iPSC clones were analyzed. The band corresponding to H3K27me3 in mutant TTF has the same intensity of that from a mutant iPSC clone for which mass-spectrometry did not detect the presence of H3K27me3. D. Quantification of AP-positive primary iPSC colonies obtained from infection of 2.5×103 Cdkn2a−/− TTF, respectively proficient (grey bar) or deficient (purple) for Ezh2, assessed in three independent experiments.