Figure 6: The conversion is dependent on Tet1.
From: Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons
(a–d) MRC5 cells reprogrammed with ANLmp and shRNA against scrambled sequence (Con_shRNA) (a), Tet1 (b), Tet2 (c) or Tet3 (d) were stained for Tuj1, TH and DAPI. (e, f) Reprogramming yield (e) and efficiency (f). *P<0.05, unpaired, two tailed Student’s t-tests versus ANLmp; n=6 wells from 3 independent experiments. (g) mRNA levels of Tet genes in MRC5 cells transduced with shRNA against control sequence, Tet1, Tet2 or Tet3. *P<0.05, unpaired, two tailed Student’s t-tests versus the preceding bar; n=6. (h) mRNA levels of p53 and Tet genes in MRC5 cells transduced with control shRNA or p53 shRNA. *P<0.05, unpaired, two-tailed Student’s t-tests versus the preceding bar; n=6. (i) mRNA levels of Tet genes in MRC5 cells with or without 10% FBS for 24 h. *P<0.05, unpaired, two-tailed Student’s t-tests versus the preceding bar; n=6. (j) MRC5 cells were treated as indicated to measure mRNA levels of Tet genes. *P<0.05 versus the first bar for each Tet gene; #P<0.05 versus the second or third bar for each Tet gene; **P<0.05 versus all other bars for each Tet gene; n=6, all with unpaired, two-tailed Student’s t-tests. (k) mRNA levels of Tet genes in ANLmp-infected MRC5 cells reprogrammed in basal media or full media for 2 days; n=6. (l-o) MRC5 cells reprogrammed with ANLmp (l) or ANLmp plus Tet1 (m), Tet2 (n) or Tet3 (o) were stained for Tuj1, TH and DAPI. (p, q) Reprogramming yield (p) and efficiency (q). *P<0.05, unpaired, two-tailed Student’s t-tests versus ANLmp for the same category; n=6. (r) 5hmC dot blot of genomic DNA from MRC5 cells reprogrammed as indicated for 2 days. Blots from 200 ng DNA were quantified. *P<0.05, unpaired, two-tailed Student’s t-tests versus ANLmp alone; n=3 independent experiments. Scale bar, 100 μm.
