DEV02422 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. The knock-in strategy employed to generate the Mox1-cre mice. The targeting vector is shown in the second line, and targeted alleles are depicted in the third (with pgk-neo cassette) and fourth lines (without pgk-cassette). The Cre-coding sequence and floxed neo cassette were replaced with part of exon 1 and intron 1 of the Mox1 gene. The pgk-neo cassette was excised by crossing with CAG-Cre mice (Sakai and Miyazaki, 1997).

• Supplemental Figure 2 -

  Fig. S2. The Epha4 enhancer activity was examined by in situ hybridization with lacZ probe for transgene (A-C) and was compared with the endogenous Epha4 expression (D-F). Embryo samples were prepared at 8.75 (A,D), 9.5 (B,E) and 11.5 (C,F) dpc from the ICR mice crossed with the permanent transgenic line established by injecting a lacZ reporter containing the 630 bp HindIII fragment.

• Supplemental Figure 3 -

  Fig. S3.The binding affinities of Mesp2/E47 heterodimer and other bHLH proteins to the E3 site. EMSA analyses were performed using the E3 probe and nuclear extracts from NIH3T3 cells transfected with expression vectors for Mesp1, Mesp2, E47, paraxis, Myod1 and twist proteins. (A) Both Mesp1 and Mesp2 bind to E3 probe in the presence of E47. (B) Paraxis does not bind to E3 probe even in the presence of E47. (C) Paraxis/E47, Myod1/E47 and twist/E47 heterodimers were not able to bind to the E3 sequence. The bands *a may correspond to E47 homodimer. The weak bands *b may be either background or Myod1 and twist homodimers.