Figure 5: TET2 is necessary for the maintenance of HOXA activity.
From: Hydroxylation of 5-methylcytosine by TET2 maintains the active state of the mammalian HOXA cluster
(a) qRT–PCR expression analysis of the three TET genes after RA treatment for 1, 3, 6 and 10 days. The left panel shows the expression levels of TET1 (black bars), TET2 (grey bars) and TET3 (green bars) as a fraction of the β-Actin expression levels. The right panel shows the expression changes of the three TET messenger RNAs during RA treatment relative to the untreated control (Cont.), internally normalized to the corresponding expression levels of Lamin-b and β-Actin. (b) qRT–PCR expression analysis of TET1 (black bars), TET2 (grey bars) and TET3 (green bars) after RA treatment and successive depletion with specific siRNA pools in single knockdowns (kd T1, kd T2 and kd T3) or in a TET1/TET2 double knockdown (kd T1/T2). NT2 cells were treated for 3 days with RA, replated and transfected with siRNAs. After 3 more days, cells were harvested and total RNA for qRT–PCR analysis was isolated. The y-axis values indicate fold change compared with the scrambled knockdown control (non-targeting pool). qRT-PCR values were internally normalized to the corresponding lamin-b and β-actin expression levels. (c) qRT–PCR expression analysis of HOXA1 (blue bars), HOXA2 (red bars), HOXA3 (green bars), HOXA4 (purple bars) and HOXA5 (light blue bars) after RA treatment and successive TET depletion with specific siRNAs in single knockdowns (kd T1, kd T2 and kd T3) or in TET1/TET2 double combination (kd T1/T2). The y-axis values indicate fold change compared with the scrambled knockdown control (non-targeting pool). qRT-PCR values were internally normalized to the corresponding lamin-b and β-actin expression levels. Treatments and measurements in (a)–(c) were repeated at least three times. s.d.'s are indicated by error bars.
