Fig. 4

SMYD3 physically associates with the NuRD complex. a, b Whole cell lysates from LM3 and SK-HEP-1 cells were prepared, and co-IP was performed using antibodies against SMYD3. Immunocomplexes were then IB tested using antibodies against the indicated proteins. IgG served as the negative control. c, d Association of SMYD3 with the NuRD complex in LM3 and SK-HEP-1 cells. Whole cell lysates were immunoprecipitated using antibodies against the indicated proteins. Immunocomplexes were then IB tested using antibodies against the indicated proteins. e Coomassie blue staining of GST-SMYD3 purified from bacterial. f GST pull-down experiments using bacterially expressed GST-fused proteins and in vitro transcribed/translated components of the NuRD complex. g Cellular extracts were obtained from LM3 cells stably expressing FLAG-SMYD3 and were immunoprecipitated with anti-FLAG antibody. The immunoprecipitates (IPs) were incubated with bulk histones and histone deacetylation (HDAC) assay buffer. The reaction mixtures were analyzed by western blotting using antibodies against the indicated histone marks or proteins