1. by Ian M. Morison, Michael R. Eccles, and Anthony E. Reeve
Imprinting of insulin-like growth factor 2 is modulated during hematopoiesis
by Ian M. Morison, Michael R. Eccles, and Anthony E. Reeve
Blood
Volume 96(9):
November 1, 2000
©2000 by American Society of Hematology

2. Diagram of exons 5 to 9 of IGF-2showing primers encompassing theApaI/AvaII polymorphic site and the resultant PCR products.Exons 7 to 9 are used by all 4 promoters, whereas exon 5 is specific to the P3-derived transcript.
Diagram of exons 5 to 9 of IGF-2showing primers encompassing theApaI/AvaII polymorphic site and the resultant PCR products.Exons 7 to 9 are used by all 4 promoters, whereas exon 5 is specific to the P3-derived transcript.
Ian M. Morison et al. Blood 2000;96:
©2000 by American Society of Hematology

3. Exon 8 and 9 IGF-2 RT-PCR products (primers 68 and 69) from 5 representative heterozygous bone marrow RNA samples digested with ApaI orAvaII.All show biallelic transcription of IGF-2.
Exon 8 and 9 IGF-2 RT-PCR products (primers 68 and 69) from 5 representative heterozygous bone marrow RNA samples digested with ApaI orAvaII.All show biallelic transcription of IGF-2. The use of bothApaI (A) and AvaII (B) excludes misinterpretation from partial enzyme digestion, whereas the product size excludes DNA contamination (Figure 1). Lane M shows λDNA digested withHindIII and EcoRI. All gels shown contain 2% agarose gel with ethidium bromide.
Ian M. Morison et al. Blood 2000;96:
©2000 by American Society of Hematology

4. Promoter 3-specific IGF-2 RT-PCR products (primers ) from 4 heterozygous normal bone marrow RNA samples.Digestion with either ApaI (A) or AvaII (B) confirms the presence of 2 transcribed alleles.
Promoter 3-specific IGF-2 RT-PCR products (primers ) from 4 heterozygous normal bone marrow RNA samples.Digestion with either ApaI (A) or AvaII (B) confirms the presence of 2 transcribed alleles.
Ian M. Morison et al. Blood 2000;96:
©2000 by American Society of Hematology

5. IGF-2 PCR and RT-PCR products (primers 68 and 69) from DNA, peripheral blood RNA, and bone marrow RNA from each of 3 healthy volunteers.Volunteers X177 and X178 express only the short allele in peripheral blood, whereas volunteer X166 expresses the long all...
IGF-2 PCR and RT-PCR products (primers 68 and 69) from DNA, peripheral blood RNA, and bone marrow RNA from each of 3 healthy volunteers.Volunteers X177 and X178 express only the short allele in peripheral blood, whereas volunteer X166 expresses the long allele. A small amount of nonspecific PCR product is present in the peripheral blood lane of volunteer X177.
Ian M. Morison et al. Blood 2000;96:
©2000 by American Society of Hematology

6. Cord blood–derived IGF-2 RT-PCR products digested with ApaI andAvaII
Cord blood–derived IGF-2 RT-PCR products digested with ApaI andAvaII.When digested with ApaI (A) or AvaII (B) they show monoallelic expression in specimens X254 and X256 but partial biallelic expression in X263 (arrows).
Cord blood–derived IGF-2 RT-PCR products digested with ApaI andAvaII.When digested with ApaI (A) or AvaII (B) they show monoallelic expression in specimens X254 and X256 but partial biallelic expression in X263 (arrows). Sample X263 also shows some shorter nonspecific (ns) PCR products that migrated in a similar position (below the bands of interest) before enzyme digestion. The ratio of the digested band intensity reflects the effect of heteroduplex formation on the digestion of PCR products. Results are consistent with a starting mix of A and B alleles of approximately 0.8:0.2. Assuming complete heteroduplex formation at the end of PCR cycling, the PCR products consist of a mixture of AA, AB, and BB duplexes in a ratio of a2:2ab:b2—ie, 0.64:0.32:0.04. On incubation with ApaI, only the BB homodimers are digested, resulting in a band ratio of 0.96:0.04 = 24:1 (undigested:digested), whereas incubation with the reciprocal enzyme AvaII products will only digest AA homodimers, resulting in a 0.36:0.64 ratio = 0.56:1. These ratios (24:1 and 0.56:1) are similar to those seen in Figure 5A and B, respectively.
Ian M. Morison et al. Blood 2000;96:
©2000 by American Society of Hematology

7. SNRPN transcription.(A) Diagram of exons 2 and 3 of SNRPN showing polymorphicBstUI site and flanking primers.
SNRPN transcription.(A) Diagram of exons 2 and 3 of SNRPN showing polymorphicBstUI site and flanking primers. (B) SNRPN RT-PCR products from 5 heterozygous bone marrow samples after digestion withBstUI. In each case imprinted transcription ofSNRPN is demonstrated. Lane M shows pUC DNA digested with MspI.
Ian M. Morison et al. Blood 2000;96:
©2000 by American Society of Hematology