Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene
J. Clin. Invest. Xueping Qu, et al. 112:1809 doi:10.1172/JCI20039 [
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Figure 1Targeted disruption of
beclin 1 in mice. (
a) Restriction maps of the wild-type
beclin 1 allele (top), the
beclin 1 targeting vector (middle), and the predicted targeted
beclin 1 allele (bottom). Restriction sites are as follows: EcoRI (E), HindIII (H). The targeting construct contains a cassette with the neomycin resistance gene (Neo) that has replaced exons 1 and 2 of the
beclin 1 gene. “X” denotes regions of homologous recombination between the targeting vector and wild-type allele. The
beclin 1 genomic fragment used as a 3′ external probe for Southern blot analysis is indicated by a solid black box. Expected sizes of the EcoRI fragments that hybridize with the probe are indicated. (
b) Southern blot analysis of genomic DNA from
beclin 1+/+ and
beclin 1+/– ES cells and mouse tails. The DNA was digested with EcoRI and hybridized with the probe indicated in
a. The sizes of wild-type (WT) and disrupted (KO) alleles are shown. (
c) PCR genotyping of genomic DNA from
beclin 1+/– and
beclin 1+/+ ES cells and mouse tail DNA. Primers 1 and 2 in
a were used to detect the wild-type allele, and primers 1 and 3 in
a were used to detect the knockout allele. (
d) Western blot analysis of Beclin 1 protein expression in
beclin 1+/+ and
beclin 1+/– ES cells and mouse lung samples. Sizes of Beclin 1 isoforms and an actin control are indicated on the right. Lung lysates were prepared from 2-month-old mice. Similar results were observed for samples from six different mice of each genotype.
