Figure 1

Math1-Cre deletes the conditional RBP-J and Patched1 alleles with high efficiency. (A) Floxing analysis of the RBP-J allele by RT-PCR with primers flanking the excised region. Samples were isolated GNPs of P7 RBP-J^lox/lox mice (pool of 4) and RBP-J^lox/lox;Math1-Cre mice (pool of 3). Bands resulting from RT-PCR show high efficiency of floxing in mutants (lane 1, 250 bp) compared to controls (lane 2, 498 bp). Lane 3 shows the negative control for the PCR. (B) quantitative real-time PCR shows decreased RBP-J mRNA levels (relative to GAPDH) in RBP-J^lox/lox;Math1-Cre mice (pool of 3) compared to RBP-J^lox/lox mice (pool of 4) (statistical analysis on means of three technical replicates for two aliquots of each cDNA pool, P = 0.0194). (C) in situ hybridisation shows loss of RBP-J mRNA in the EGL of E18.5 RBP-J^lox/lox;Math1-Cre embryos. Images were taken at the same magnification; scale bar represents 200 μm. (D) Floxing analysis of the Ptc1 allele by RT-PCR with primers flanking the excised region. Samples were isolated GNPs of P7 Ptc1^lox/lox mice (pool of 4) and Ptc1^lox/lox;Math1-Cre mice (pool of 3). Bands resulting from RT-PCR confirm high efficiency of floxing in mutants (lane 1, 250 bp) compared to controls (lane 2, 450 bp). Lane 3 shows the negative control for the PCR. (E) Quantitative real-time PCR indicates increased Gli1 mRNA levels (relative to GAPDH) in Ptc1^lox/lox;Math1-Cre mice (pool of 3) compared to Ptc1^lox/lox mice (pool of 4), supporting activation of the Hh pathway by Ptc1 deletion (statistical analysis on means of three technical replicates for two aliquots of each cDNA pool, P = 0.0132).