Figure 2
Inhibition of H3K27me3 activity impairs mesenchymal to epithelial transition during iPSC generation.
(a) Western blot analysis using anti Ezh2, anti-H3K27Me3 and control anti-pan H3 antibodies in hFibs treated with H3K27me3 inhibitor GSK-126. Uncropped/Full blots images are provided in Supplementary Fig. 6 a-c. (b) Relative expression of p53 and p21 mRNA in DMSO control and GSK treated hFibs. (c) Representative bright field and β- gal staining images upon treatment of hFibs with DMSO and GSK-126. (d) Western blot analysis using anti Ezh2, anti-H3K27Me3 and control anti-pan H3 antibodies in H9 hESC treated with DMSO or H3K27me3 inhibitor GSK-126. Uncropped/Full blots images are provided in Supplementary Fig. 6 d–f. (e) Tra 1-60 DAB staining in IMR-90 iPSC, H9 hESC cultures treated with DMSO and GSK-126. (f) Schematic representation of reprogramming protocol and time points of addition of GSK to the reprogramming cultures. (g) Total number of reprogrammed colonies generated three weeks post OSKM transduced hFibs and upon GSK treatment at indicated time points in schema (h) Tra 1-60 DAB staining on day 21 post addition of reprogramming factors and GSK treatment (i) E-cad DAB staining on day 10 post addition of reprogramming factors and GSK treatment as indicated in schema.
