Extended Data Fig. 3: NSD2 and H3K36me2 expression in patient tumors and prostate cancer cell lines.

a) Representative immunofluorescence (IF) images of NSD2 in benign prostate, primary prostate cancer (PCa), and metastatic CRPC tissue microarray.Scale bar:50 µm. b) Representative multiplex IF images of NSD2 and CK-8 in adjacent benign and primary prostate cancer lesions in patient prostatectomies (n = 5 biological replicates, Scale bar:50 µm). c) Integrated optical density quantification of NSD2 IF staining in benign (n = 10), primary PCa (n = 10), and mCRPC (n = 5) tissues. Box plot center, median; box, quartiles 1-3; whiskers, min and max values. d) Box plots of normalized ChIP-seq reads of distinct activating and repressive histone modifications at NSD2-dependent and NSD2-independent AR sites in VCaP cells (n = top 2000 sites, two-sided t-test). Box plot center, median; box, quartiles 1-3; whiskers, quartiles 1-3 ± 1.5 × interquartile range; dot, outliers. e) Box plots of normalized ChIP-seq reads of NSD2-catalyzed H3K36me2 and EZH2/PRC2-catalyzed H3K27me3 histone marks at NSD2-dependent and NSD2-independent AR sites in LNCaP cells (n = top 2000 sites, two-sided t-test). Box plot center, median; box, quartiles 1-3; whiskers, quartiles 1-3 ± 1.5 × interquartile range; dot, outliers. f) ChIP-seq read-density tracks of histone modification within a Chr10 locus in VCaP cells. NSD2-dependent and independent AR sites are marked in the tracks below with representative enhancers highlighted. g) ChIP-seq read-density box plots showing H3K36m2 (top) and H3K27me3 (bottom) signals at AR sites in NSD2-KO or WT LNCaP cells (n = top 2000 sites, two-sided t-test). Box plot center, median; box, quartiles 1-3; whiskers, quartiles 1-3 ± 1.5 × interquartile range; dot, outliers. h) ChIP-seq read-density tracks of H3K36me2 and H3K27me3 within a Chr10 locus in NSD2 WT and KO LNCaP cell lines. NSD2-dependent and independent AR sites are marked and highlighted.

Source data