1. A novel mutation of HFE explains the classical phenotype of genetic hemochromatosis in a C282Y heterozygote 
Daniel F. Wallace, James S. Dooley, Ann P. Walker 
Gastroenterology 
Volume 116, Issue 6, Pages (June 1999)
DOI: /S (99)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Family tree. Patient RFH is indicated by an arrow; his sister has an identical genotype and evidence of iron overload. One daughter is heterozygous for C282Y and the other is heterozygous for IVS3 + 1G → T, indicating that the two mutations occur on different HFE alleles. Percent transferrin saturations and serum ferritin concentrations (μg/L) are shown where available. HII, hepatic iron index (μmol · g−1 · yr−1);Vx, amount of iron venesected to normalize serum ferritin.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

3. Fig. 2
The IVS3 + 1G → T mutation functionally alters mRNA splicing. (A and B) Southern blots of allele-specific RT-PCRs hybridized with an exon 4 probe, amplified from (A) 282C allele or from (B) 282Y allele. Lane 1, patient RFH, primary leukocytes; lane 2, patient RFH, Epstein–Barr virus–transformed lymphoblastoid cell line; lane 3, control leukocyte, homozygous for H63D; lane 4, control duodenum, wild-type sequence; lane 5, control fibroblast, heterozygous for C282Y; lane 6, no reverse-transcription control; lane 7, no template control; lane 8, genomic DNA control. The bands corresponding to the cognate splicing of exons (717 base pairs) and alternative splicing of exons 2-4 (441 base pairs) are indicated. The 282Y-specific primer gave amplification of both the cognate and alternatively spliced products only in samples containing C282Y. The 282C primer amplified only the alternatively spliced product (exons 2-4) in patient RFH, whereas the cognate product (exons 2-3-4) was also amplified from all other RNA samples. (C) Schematic representation of HFE mRNA splicing around exon 3. Wild-type C282Y and H63D alleles produced 2 splice variants. (i) Splicing of exons in the normal manner; (ii) skipping of exon 3. *The IVS3 + 1G → T mutation resulted in complete skipping of exon 3, consistent with the invariance of this nucleotide in functional splice sites.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

4. Fig. 2
The IVS3 + 1G → T mutation functionally alters mRNA splicing. (A and B) Southern blots of allele-specific RT-PCRs hybridized with an exon 4 probe, amplified from (A) 282C allele or from (B) 282Y allele. Lane 1, patient RFH, primary leukocytes; lane 2, patient RFH, Epstein–Barr virus–transformed lymphoblastoid cell line; lane 3, control leukocyte, homozygous for H63D; lane 4, control duodenum, wild-type sequence; lane 5, control fibroblast, heterozygous for C282Y; lane 6, no reverse-transcription control; lane 7, no template control; lane 8, genomic DNA control. The bands corresponding to the cognate splicing of exons (717 base pairs) and alternative splicing of exons 2-4 (441 base pairs) are indicated. The 282Y-specific primer gave amplification of both the cognate and alternatively spliced products only in samples containing C282Y. The 282C primer amplified only the alternatively spliced product (exons 2-4) in patient RFH, whereas the cognate product (exons 2-3-4) was also amplified from all other RNA samples. (C) Schematic representation of HFE mRNA splicing around exon 3. Wild-type C282Y and H63D alleles produced 2 splice variants. (i) Splicing of exons in the normal manner; (ii) skipping of exon 3. *The IVS3 + 1G → T mutation resulted in complete skipping of exon 3, consistent with the invariance of this nucleotide in functional splice sites.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

5. Fig. 2
The IVS3 + 1G → T mutation functionally alters mRNA splicing. (A and B) Southern blots of allele-specific RT-PCRs hybridized with an exon 4 probe, amplified from (A) 282C allele or from (B) 282Y allele. Lane 1, patient RFH, primary leukocytes; lane 2, patient RFH, Epstein–Barr virus–transformed lymphoblastoid cell line; lane 3, control leukocyte, homozygous for H63D; lane 4, control duodenum, wild-type sequence; lane 5, control fibroblast, heterozygous for C282Y; lane 6, no reverse-transcription control; lane 7, no template control; lane 8, genomic DNA control. The bands corresponding to the cognate splicing of exons (717 base pairs) and alternative splicing of exons 2-4 (441 base pairs) are indicated. The 282Y-specific primer gave amplification of both the cognate and alternatively spliced products only in samples containing C282Y. The 282C primer amplified only the alternatively spliced product (exons 2-4) in patient RFH, whereas the cognate product (exons 2-3-4) was also amplified from all other RNA samples. (C) Schematic representation of HFE mRNA splicing around exon 3. Wild-type C282Y and H63D alleles produced 2 splice variants. (i) Splicing of exons in the normal manner; (ii) skipping of exon 3. *The IVS3 + 1G → T mutation resulted in complete skipping of exon 3, consistent with the invariance of this nucleotide in functional splice sites.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions