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  Indian J Med Microbiol
 

Figure 1: Schematic illustration of the Zeb1 flox allele and deletion of exon 6. (a) Gray boxes indicate exons, and triangles indicate the location of the loxP sites flanking exon 6. The location of F1, F2, R1, and R2 genotyping primers are shown by arrows. Cre recombinase activity leads to excision of exon 6 along with R1 and F2 primer sites. (b) The loxP sites were confirmed via genotyping with pair of primers; F1-R1 for proximal site and F2-R2 for distal site. Seven days after intracameral 4-OHT (+) or vehicle (−) injection, the endothelium was isolated and genomic DNA was purified for PCR genotyping. Exon 6 of Zeb1 was deleted in the corneal endothelium following intracameral 4-OHT injection in Zeb1flox/flox: UBC-CreERT2 mice but not in the other mice. In Zeb1flox/flox: UBC-CreERT2 mice that received intracameral 4-OHT injection, PCR genotyping with F1-R2 primer pair generated the expected 367 bp product while F1-R1 and F2-R2 reactions did not generate any products due to the loss of R2 and F1 sites following Cre-mediated recombination. In all other groups, F1-R1 and F2-R2 reactions generated the expected 295 bp and 512 bp products respectively while F1-R2 reaction did not generate the expected 367 bp product, indicating Zeb1 was not altered

Figure 1: Schematic illustration of the Zeb1 flox allele and deletion of exon 6. (a) Gray boxes indicate exons, and triangles indicate the location of the loxP sites flanking exon 6. The location of F1, F2, R1, and R2 genotyping primers are shown by arrows. Cre recombinase activity leads to excision of exon 6 along with R1 and F2 primer sites. (b) The <i>loxP</i> sites were confirmed via genotyping with pair of primers; F1-R1 for proximal site and F2-R2 for distal site. Seven days after intracameral 4-OHT (+) or vehicle (−) injection, the endothelium was isolated and genomic DNA was purified for PCR genotyping. Exon 6 of <i>Zeb1</i> was deleted in the corneal endothelium following intracameral 4-OHT injection in Zeb1<sup>flox/flox</sup>: UBC-CreERT2 mice but not in the other mice. In Zeb1<sup>flox/flox</sup>: UBC-CreERT2 mice that received intracameral 4-OHT injection, PCR genotyping with F1-R2 primer pair generated the expected 367 bp product while F1-R1 and F2-R2 reactions did not generate any products due to the loss of R2 and F1 sites following Cre-mediated recombination. In all other groups, F1-R1 and F2-R2 reactions generated the expected 295 bp and 512 bp products respectively while F1-R2 reaction did not generate the expected 367 bp product, indicating <i>Zeb1</i> was not altered