Reassessment of the algorithm for prediction of liver fibrosis in patients with features of the metabolic syndrome

✍ Scribed by Fabrice Lainé; Claude Bendavid; Jeff Morcet; Michéle Perrin; Catherine Massart; Yves Deugnier

• Publisher: John Wiley and Sons
• Year: 2006
• Tongue: English
• Weight: 70 KB
• Volume: 43
• Category: Article
• ISSN: 0270-9139
• DOI: 10.1002/hep.21038

✦ Synopsis

Mutational analysis of the target oligonucleotide used for the EMSAs revealed that all mutations in the canonical NFB binding site and not the flanking DNA affected the binding of NFB (the upper two bands on our EMSAs), In contrast, only the most 3Ј mutations in the canonical NFB binding sequence and some flanking DNA in the oligonucleotide affected binding of the lower band. These results, combined with the fact that the lower band on the gel did not supershift with any of the NFB antibodies used, make it unlikely that the lower band contains NFB. Finally, we were able to immunodeplete the protein responsible for the lower band using anti-nucleolin antibodies.

Given these results, we were left to conclude that the only protein species accounting for significant NFB binding in the fetal liver extracts was nucleolin. Within the context of our studies, this was important because it supported the conclusion that NFB is not active in late gestation fetal liver. We hope that our comments are adequate to address Dr. Bernstein's concerns.