1. Volume 135, Issue 1, Pages 282-291.e1 (July 2008)
The Kruppel-Like Factor 6 Genotype Is Associated With Fibrosis in Nonalcoholic Fatty Liver Disease 
Luca Miele, Gary Beale, Gillian Patman, Valerio Nobili, Julian Leathart, Antonio Grieco, Marilena Abate, Scott L. Friedman, Goutham Narla, Elisabetta Bugianesi, Christopher P. Day, Helen L. Reeves 
Gastroenterology 
Volume 135, Issue 1, Pages e1 (July 2008)
DOI: /j.gastro
Copyright © 2008 AGA Institute Terms and Conditions

2. Figure 1
Increased wt KLF6 expression with advanced stages of NAFLD. Either Total KLF6 mRNA isoforms (represented by black bars) or the wt isoform only (grey bars) have been quantified by semiquantitative TaqMan RT-PCR. The individual Brunt et al25 histology scores and KLF6 RQ values are shown in Table 1, whereas the mean expression levels (RQ log) ± standard error within histologically defined grades of NAFLD are shown in this Figure (n = 31; P values are shown). (A) Stepwise increase in both Total KLF6 isoforms and wt KLF6 increased stepwise in association with increasing levels of inflammation (A). Although wt KLF6 also was increased significantly in association with advanced steatosis (B) and fibrosis (C), Total KLF6 isoforms were not. (A) ■, Total, P = .001; , wt, P = .010; (B) ■, P = .049; , P = .001; (C) ■, NS; , P = .03.
Gastroenterology  , e1DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

3. Figure 2
KLF6 IVS1-27G>A promotes alternative splicing of KLF6 and abrogates the up-regulation of αSMA and collagen 1. Cells were transfected with either transfection reagent alone (con), a control minigene expressing LacZ (lacz), a KLF6 minigene with the IVS1-27G>A SNP (G>A), or a KLF6 wt minigene construct (wt). (A) Agarose gel electrophoresis of cDNA products amplified using 5′ and 3′ flanking primers detecting all KLF6 isoforms identified is shown. KLF6 in the wt minigene transfected cells was predominantly the full-length wt isoform, whereas in the cells transfected with the KLF6 IVS1-27G>A minigene there were additional smaller bands representing alternative splice forms. Relative quantities of these isoforms were confirmed by semiquantitative RT-PCR (n = 3, data not shown). (B) Semiquantitative RT-PCR data from RNA extracted from stably transfected LX-2 cells harvested at 1 week. Total KLF6 was increased in cells transfected with either of the KLF6 minigene constructs, whereas only a relatively modest amount of this total KLF6 was the wt isoform in the G>A–transfected cells. (C) The marked increase in wt KLF6 in the LX-2 cells transfected with the wt KLF6 minigene was associated with significant increases in both αSMA and collagen I mRNA, which was not present in those cells transfected with the G>A SNP containing minigene-expressing dominant-negative KLF6 isoforms in addition to the wt isoform.
Gastroenterology  , e1DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions