Figure 5.

NSD1 is essential for DMAPT-induced BIM and p21 expression. (a) BIM and p21 levels in untreated and DMAPT-treated UMUC-3 cells additionally transfected with siRNA against luciferase as a negative control, NSD1, SETD2, or both NSD1 and SETD2. Cells were treated after 3 days of siRNA transfection with DMAPT and harvested for proteins after 24 h of DMAPT treatment. NSD1 siRNA prevented DMAPT-induced BIM and p21. For unknown reasons, basal p21 levels in NSD1 siRNA-treated cells showed remarkable experimental variability in UMUC-3 cells. Densitometry values show comparison between untreated controls and DMAPT-treated cells with respective controls normalized to 1. (b) NSD1 and SETD2 mRNA levels in siRNA-transfected cells. Respective mRNAs were measured 4 days after siRNA transfection. (c) SETD2 is required for DMAPT-mediated growth inhibition. Cells were plated in a 96-well plate and treated with DMAPT for 48 h. Cell proliferation was measured using BrDU-incorportation ELISA