Supplementary Figure 8: Generation of the Stoml3^lacZ and Stoml3^StrepII mouse strains

(a) Schematic representation of the targeting vector, the wild type Stoml3 locus, and the mutated Stoml3^lacZ allele, before and after removal of the self-excision neomycin (cre, neo) cassette. A 12 kb genomic region of Stoml3 locus containing exon 1 (E1, black), NLS-lacZ (blue), DTA (yellow), the self-excision neomycin cassette, loxP (red arrowhead), and SpeI (S) restriction sites are depicted. Green lines indicate the predicted fragment sizes obtained after SpeI digestion of genomic DNA. A green bar shows the 5’ sequence used as a probe for Southern blot analyses shown in b. Blue lines indicate the predicted fragment sizes obtained by genotyping the tail genomic DNA as shown in c. (b) Southern blot analysis of SpeI digested tail genomic DNA from Stoml3^+/lacZ and wild type mice. (c) Genotyping analysis of tail genomic DNA from Stoml3^+/lacZ, wild type, and Stoml3^lacZ/lacZ mice. Stoml3-LacZ F and LacZ int R primers amplified a 649 bp fragment from the Stoml3^lacZ mutant allele. Stoml3-LacZ F and Stoml3-LacZ R primers amplified a 875 bp fragment from the Stoml3 wild type allele, M: 100 bp ladder (Invitrogen). (d) Schematic representation of the targeting vector, the wild-type Stoml3 locus, and the mutated Stoml3^StrepII allele, before and after removal of the neomycin (neo) cassette. A 12 kb genomic region of Stoml3 locus containing exon 1 (E1, black), Strep-TagII (red), DTA (yellow), the neomycin cassette (neo), loxP (blue arrowhead), and SpeI (S) restriction sites are depicted. Green lines indicate the predicted fragment sizes obtained after SpeI digestion of genomic DNA. A green bar shows the 5’ sequence used as a probe for Southern blot analyses shown in e. Blue lines indicate the predicted fragment sizes obtained by genotyping the tail genomic DNA as shown in f. (e) Southern blot analysis of SpeI digested tail genomic DNA from wild type, Stoml3^+/StrepII and Stoml3^{StrepII/StrepII} mice. (f) Genotyping analysis of tail genomic DNA from wild type, Stoml3^+/StrepII, and Stoml3^{StrepII/StrepII} mice. Stoml3-Strep F and Stoml3-Strep R2 primers amplified a 321 bp fragment from the Stoml3^StrepII mutant allele. Stoml3-Strep F and Stoml3-Strep R1 primers amplified a 438 bp fragment from the Stoml3 wild type allele, M: 100 bp ladder (Invitrogen).