DEV01179 Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. GFP expression in transgenic mouse embryos. (A-L) Transient transgenic mouse embryos containing the M250+DNheI:GFP construct were isolated at E11.5 and stained with antibodies. Sections of the ventral portion of the neural tube are shown. (A-C) GFP labeling is strongest in Hb9⁺ motoneurons. GFP is also detected at lower levels in cells dorsal to the motor column, and in dorsal root ganglion cells adjacent to the neural tube. (D-F) Likewise, GFP labeling is strongest in Isl1⁺ motoneurons. (G-I) Lhx3 marks a subset of medial motoneurons and V2 interneurons. Strong GFP labeling is detected in the medial motor column, but very few of the V2 interneurons appear to label with GFP. (J-L) Likewise, Chx10 labels V2 interneurons, however, very few of the cells appear to express GFP.

• Supplemental Figure 2 -

  Fig. S2. bHLH factor activation of Hb9. (A) Luciferase assays with wild type (black) and E box mutant (white) derivatives of M250 transfected into CV1 cells with the indicated bHLH expression constructs. The activation of the reporter is dependent upon functional E box elements, and is stimulated by both Ngn2 and NeuroM, particularly when E47 is overexpressed. (B) Activation of the M250 reporter in 293 cells by Ngn2 and NeuroM (NeuM) is mediated by direct DNA binding of these factors. AQ point mutations in the DNA binding domains of these factors (Lee and Pfaff, 2003) block their ability to activate the reporter. (C) Dimers of Ngn2:E47, NeuroM:E47, and NeuroD:E47 bind to the E box elements within the M50 and M100 regions of Hb9.