Figure 4

Methylation profile and antisense transcript analysis for the novel DMR identified in intron 11 of M. domestica IGF2R. ( A ) IGF2R is represented graphically. The three sets of primers used in strand-specific RT-PCR analysis are labeled as 1, 2 and 3. The reverse transcription primers for anti-sense strand detection are labeled as 1a, 2a, and 3a while those for sense-strand detection are labeled as 1b, 2b, and 3b. ( B ) Parent-of-origin analysis was performed at the DMR by analyzing methylation of cloned alleles for an individual bearing an (A/T) SNP at chr2: 442,443,695 with a mother homozygous for A at the same location. Only the allele inherited from the mother is methylated. ( C ) Strand-specific RT-PCR analysis using primers described above shows a PCR product only by reverse transcription utilizing 1b, 2b band 3b; L represents the 100 bp DNA ladder. This demonstrates that the gene is expressed at this locus but there is no associated anti-sense transcript.