Figure 3

Impact of Tet1 and Tet2 knockdown on adipogenesis via 5-hmC-mediated Pparg regulation. The amount of triglycerides measured by Oil-Red-O staining in sextuplets upon Tet1 or Tet2 knockdown followed by the induction of adipogenesis (a). Tet1 and Tet2 knockdown efficiency based on Tet1 and Tet2 mRNA levels measured by qPCR in triplicates (b, c). Pparγ and Fabp4 expression levels, measured by qPCR in triplicates, in adipogenic induced 3T3-L1 cells after Tet1 and Tet2 knockdown (d, e). Global 5-hmC level during differentiation measured by GC–MS (f) and a locus-specific 5-mC and 5-hmC at the Pparγ locus quantified by MeDIP (g) and hMeDIP (h), respectively, in duplicates upon Tet1 or Tet2 knockdown followed by the induction of adipogenesis. The data were analyzed by Student's t-test, and *P<0.05 and **P<0.01 indicate significance between the knockdown versus the control; ‘ns’ presents ‘not significant’; d0, d3, d6, d9, d12 indicate days after adipogenesis induction.