Figure 7
Impaired neural differentiation of TET1-deficient hESCs is accompanied by decreased 5hmC content in PAX6 promoter. (a,b) qRT-PCR measurement of the expression levels of the indicated neural lineage markers in cells differentiated from wild-type (WT) or TET1-deficient (CDKO-1 and CDKO-2) H9 hESCs under the morphogen-free condition at day 14 (a) and day 7 (b). *p < 0.05, paired Student’s t-test, n = 3. (c–e) The two TET1 mutant hESC lines (CDKO1 and CDKO2) infected with lentiviruses expressing GFP or FLAG-TET1CD were differentiated in the morphogen-free condition. Embryoid bodies at day 10 of differentiation were analyzed by qRT-PCR for the expression levels the neuroecdoderm genes PAX6 (c), FOXG1 (d) and SOX1 (e). *p < 0.05, paired Student’s t-test, n = 6. (f,g) hMeDIP measurement of 5hmC content at the promoter region of PAX6, FOXG1 or miR218, a gene unrelated to early neural differentiation, in EBs formed under morphogen-free condition (f) or in undifferentiated hESCs (g). *p < 0.05, Student’s t-test, n = 3. (h,i) MeDIP measurement of 5mC content at the promoter of PAX6, FOXG1 or miR218 in EBs formed under morphogen-free condition (h) or in undifferentiated hESCs (i).
