Figure 3.

MMSET interacts with PCNA via its N-terminus and PCNA depletion promotes MMSET stability. (A) U2OS cells were immunoprecipitated with PCNA or mouse IgG and the resulting samples were examined for MMSET or PCNA by western blot. (B) Left: T98G cells were transfected with the indicated siRNAs and synchronized in S phase by the serum starvation and release method. Lysates were prepared 48 hours later and examined for the indicated proteins. Right: cell-cycle profiles of the samples. (C) Schematic representation of MMSET fragments used for the GST pull-down assay. (D) A GST-pull down assay was carried out between purified PCNA and GST-MMSET fragments. Coomassie blue staining shows GST fusion proteins used in the pull-down assay (middle). Data shown are representative of 3 independent experiments.