In a drug discovery project to discover compounds against methyltransferase NSD2, we needed to establish the baseline methylation of histone H3 without NSD2 expression.
To evaluate the activity of NSD2 we chose to use U-2 OS human osteosarcoma cells due to a relatively high expression of endogenous NSD2 protein. Three siRNA molecules were generated to knock down NSD2 in the cells. To validate the methyltransferase activity in the cells, Western Blot analysis was performed with antibodies specific to dimethylated histone H3 molecules. The analysis successfully showed that the reduction in NSD2 expression is directly correlated with dimethylation of lysine 36 in Histone 3 molecules.