Figure 6. Nuclear heparanase may control H3 methylation profiles by recruiting LSD1 and excluding MLL from gene promoters in T cells. (A) LSD1 ChIP assays were performed on Jurkat T cells transfected with heparanase RNAi3 or a negative siRNA control (Mock). Real-time PCR was performed using primers covering the -0.15 kb proximal promoter of the CD69, IFN-γ, IL-2 and TNF-α genes. (B) Immunoprecipitation of heparanase from cross-linked sonicated nuclear lysates of either NS or PI-treated T cells followed by immunoblot analysis with an anti-LSD1 antibody. No antibody (no Ab) was used as a negative control. (C) Jurkat T cells untreated (Ctrl) or pre-treated with 3 mM pargyline (PAR) and non-stimulated or stimulated with PI for 2 h. Total RNA was extracted and TaqMan real-time PCR performed for CD69, IFN-γ, IL-2 and TNF-α transcripts. Results are plotted as arbitrary copies relative to the corresponding NS sample and normalized to GAPDH. (D) SET7/9, (E) MLL and (F) H3K9ac ChIP assays were performed as in (A). Data represent the mean ± SE of three independent experiments. (G) Proposed model of the interplay of nuclear heparanase with LSD1, RNAP II, MLL and histone methylation marks.