NEURL1 inhibits the Notch pathway and colocalizes with Jagged1. Stable transfectants of DAOY and R300 show reduced Notch pathway activation with decreased expression of target genes (A) HES1 and (B) HEY1. In (A) and (B), DAOY: NEURL1 (n = 5), ΔRING (n = 5), and vector only (n = 5); R300: NEURL1 (n = 4), ΔRING (n = 4), and vector only (n = 4); n, number of individual stable clones. (C) Immunofluorescence microscopy of DAOY NEURL1 and ΔRING stable expressing clones transiently transfected with JAG1 shows reduced staining of Jagged1 in the NEURL1 transfectant. Arrows show colocalization of Neuralized1 and Jagged1 in punctate clusters at the cell surface. (D) Jagged1 mRNA levels are also decreased through the expression of NEURL1. DAOY cells: NEURL1 (n = 5), ΔRING (n = 5), and vector only (n = 5); R300 cells: NEURL1 (n = 4), ΔRING (n = 4), and vector only (n = 4); n, number of individual stable clones. (E) The Western blots show decreased expression of endogenous Jagged1 in NEURL1 stable transfectants compared with controls.