SETDB1 target gene expression is regulated by SETDB1 ubiquitination.

<p>HeLa cells were transfected with pcDNA3-hSETDB1 (SETDB1 WT), pcDNA3-hSETDB1 K867R (SETDB1 K867R), or pcDNA3 (Empty). (A and D) mRNA levels were normalized to those of cyclophilin A mRNA. All values are expressed as the mean ± SEM (n = 3). *<i>P</i> < 0.05 or **<i>P</i> < 0.01 compared with the value of Empty. (B) Whole cell extracts were prepared and aliquots [30 μg (for SETDB1) or 10 μg (for lamin B2) protein/lane] were subjected to SDS-PAGE. Immunoblots were probed with anti-SETDB1 antibody or anti-lamin B2 antibody. (C) Purified genomic DNA after ChIP was analyzed by real-time PCR. Data was normalized to the input DNA. Fold change was calculated relative to Empty. All values are expressed as the mean ± SEM (n = 3). **<i>P</i> < 0.01 compared with the value of Empty.