Supplementary Figure 1: Rfx7 phylogeny and conditional-knockout strategy.

(a) Percent identity matrix of full RFX7 protein sequences generated with Clustal 2.1. (b) Phylogenetic tree based on full protein alignment (Neighbor-joining tree without distance corrections, Clustal 2.1); the bar line (distance scale) indicates the percentage of variation among species, i.e. 0.1 represent 10% difference between two sequences. (c) Alignment of RFX7 DNA Binding Domain for the indicated species (JalView). The compared protein sequences correspond to the following species: Human, Homo sapiens; Chimpanzee, Pan troglodytes; Gorilla, Gorilla gorilla; Macaque, Macaca mulatta; Orangutan, Pongo pygmaeus; Mouse, Mus musculus; Rat, Rattus norvegicus; Rabbit, Oryctolagus cuniculus; Guinea pig, Cavia porcellus; Pig, Sus scrofa; Cow, Bos taurus; Chicken, Gallus gallus; Xenopus, Xenopus tropicalis; Zebrafish, Danio rerio. (d) Scheme showing the targeting strategy for the conditional knockout of the murine Rfx7 gene. The vector was designed such as the 5’ loxP site is inserted upstream of exon 3 and the target region is 1.96 kb including exons 3-4. The loxP/FRT flanked Neo cassette is inserted downstream of exon 4. The selection cassette (Neo) was excised by crossing floxed mice with FLP deleter line. The two loxP sites shown allow Cre-mediated deletion of exons 3 (Ex 3) and 4 (Ex 4). PCR primers b/c discriminate wild type (wt) and floxed (fl) Rfx7 alleles and primers a/c wild type and knockout (ko) Rfx7 alleles. Arrows on diagram indicate PCR primer positions. PCR products obtained for each genotype are shown. LA: long arm, MA: middle arm, SA: short arm, FRT: flippase recognition target, Neo: neomycine cassette