Fig. 5

Silencing of NSD2 abrogates tumorigenicity in vitro. Panels a-g show in vitro analyses of NSD2 silencing in a mouse metastatic cell line (NPK cells) and a human advanced prostate cancer cell line (DU145 cells). Cells were infected with control shRNA or two independent shRNAs for mouse or human NSD2, respectively. a, b Validation of NSD2 silencing in NPK and DU145 cells, as indicated, using quantitative real-time PCR (qPCR). c Western blot analyses of NSD2-silenced or control NPK and DU145 cells, as indicated, showing reduced expression of NSD2, which is accompanied by reduction of the H3K36me2 mark, but not the H3K36m1 or the other histone marks shown. The position of a molecular marker is shown; uncropped images are provided in Supplementary Figure 5. d, e Colony formation assays in NPK and DU145 cells, as indicated showing quantification (left) representative images (right). f, g Invasion assays in NPK and DU145 cells, as indicated showing quantification (left) and representative images (right). Experiments were done in three independent biological replicates each in triplicate; p-values were calculated using a two-tailed Student's t-test. Error bars represent the standard deviation (s.d.) from the mean