Figure 5.

PARylation of NSD2 inhibits its enzyme activity. A, schematic of the experimental strategy. NSD2 was incubated with individual cofactors or together with PARP1 for PARylation reaction as outlined in Step 1. Subsequently the reactions were terminated by adding Olaparib and incubated on ice for 30 min as in Step 2. B, one-third of the PARylation reaction was removed and analyzed by immunoblot to assay for PARylation. C, ³H-labeled SAM and purified HeLa oligonucleosomes were added for the NSD2 histone methyltransferase reaction as outlined in Step 3 and methylation activity was analyzed by a filter-binding assay that measures incorporation of ³H-labeled methyl groups. Two independent experiments are shown.