Figure 2

Reduced 5hmC level in TET1-deficient hESCs. (a,b) Dot blot (a) and quantification (b) of 5hmC in WT and TET1-deficient hESCs. *p < 0.001, paired Student’s t-test, n = 3. (c,d) Dot blot (c) and quantification of 5mC in WT and TET1-deficient hESCs. Full images of the dot blots without any imaging processing are displayed in (a) and (c). (e,f) 5hmc staining (e-e”) and quantification (f) in WT (e) and TET1-deficient (e’ and e”) hESCs. *p < 0.001, Student’s t-test, n = 5. (g,h) 5mC staining (g-g”) and quantification (h) in WT (g) and TET1-deficient (g’ g”) hESCs. Scale bar, 100 µm. (i,-k) The two TET1 mutant hESC lines (KO1 and KO2) were infected with lentivirus expressing GFP or FLAG-tagged TET1 catalytic domain (F-TET1CD). Total cell lysates were blotted with the indicated antibodies (i). Genomic DNA isolated from these cells were dot-blotted with antibodies against 5hmC (j) or 5mC (k). *p < 0.05, paired Student’s t-test, n = 4.