LOH and mutation analysis of Pten in Pten+/- and Pten+/-;Mlh1-/- mice. A) Representative photomicrographs of microdissected invasive (1) and CAH (2) lesions. B) PCR based analysis of LOH of Pten in endometrial lesions from CD-1 Pten+/- mice. Lane M, 1kb ladder. Lane 1, amplification of DNA from microdissected normal tissue from a Pten+/- mouse showing both mutant (*) and wild-type (arrow head) alleles. Lane 2, amplification of DNA from microdissected tissue from a wild-type mouse. Lane 3 and 6, amplification of DNA from two microdissected lesions without LOH. Lane 4, 5 and 7, amplification of DNA from three microdissected lesions showing LOH of the wild-type Pten allele. C) LOH at microsatellite loci D19Mit40 and D19Mit71 at Chromosome 19 in the CAH with LOH of Pten from a CD-1 Pten+/- mouse. Lane N, microdissected normal tissue showing two alleles. Lane C, microdissected CAH showing LOH at both loci tested. D) A frameshift mutation in exon 5 of Pten from a Pten+/-;Mlh1-/- lesion without LOH of Pten. A 1-bp deletion (T) at codon 146 is detected (arrow), which leads to a stop at codon 146.