DEV02658 Supplementary Material

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• Supplemental Figure 1 -

  Fig. S1. Gene disruption at the CL2L1 locus. (A) Schematic representation of Cp2l1^flox and Cp2l1^del alleles. Red and blue arrows indicate primers used for PCR detection of the modified and wild-type alleles, respectively (see B). Primers used for detection of flox and deleted alleles were: 5′-cgattcgcagcgcatcgccttct-3′ and 5′-gtagcaggagagaaccgac-3′; primers used for detection of the wild-type allele were: 5′-ccagcggtgaacgtacatg-3′ and 5′-aatccctgaggcaccgagaa-3′. (B) Genotyping of Cp2l1^flox and Cp2l1^del alleles. The Cp2l1^flox and Cp2l1^del alleles were distinguished by the red primers shown in A; the wild-type allele was amplified by the blue primers shown in A. (C) Confirmation of mRNA sequences transcribed from the Cp2l1^del allele. Above: PCR using primers that amplify the ORF of Cp2l1 mRNA. Only the deleted, not the intact, Cp2l1 transcript was generated in mice homozygous for the Cp2l1^del allele. Below: A frame-shift mutation was found in exon 2-deleted mRNA transcribed from the Cp2l1^del allele. Primers used for amplification of the ORF were: 5′-atgctgttctggcacacgca-3′ and 5′-tcagagtccacacttcagg-3′. (D) The morphology of the submandibular gland was similarly defective in mice homozygous for the Cp2l1^tra and Cp2l1^del alleles. The grandular convoluted duct (labeled G) did not develop in these mutants. Scale bars: 100 μm.