Expression and functional analysis of intestinal organic cation/l-carnitine transporter (OCTN) in Crohn's Disease Marc Girardin, Serge Dionne, Philippe.

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Expression and functional analysis of intestinal organic cation/l-carnitine transporter (OCTN) in Crohn's Disease Marc Girardin, Serge Dionne, Philippe Goyette, John Rioux, Alain Bitton, Ihsan Elimrani, Patrick Charlebois, Ijaz Qureshi, Emile Levy, Ernest G. Seidman Journal of Crohn's and Colitis Volume 6, Issue 2, Pages 189-197 (March 2012) DOI: 10.1016/j.crohns.2011.08.003 Copyright © 2011 European Crohn's and Colitis Organisation Terms and Conditions

Figure 1 Representative Western blots of intestinal OCTN1 and OCTN2. Actin was used as a standard. CD = Crohn's disease and C = control. Journal of Crohn's and Colitis 2012 6, 189-197DOI: (10.1016/j.crohns.2011.08.003) Copyright © 2011 European Crohn's and Colitis Organisation Terms and Conditions

Figure 2 Intestinal OCTN protein level determination by Western blot in inflammatory bowel disease patients and controls, using tissue actin levels as standard. A) OCTN1 protein expression in human intestinal tissue comparing ileum (n=31) and colon (n=16) (*p<0.0002). B) OCTN2 protein expression in human intestinal tissue comparing ileum (n=35) and colon (n=18). C) OCTN1 protein expression in human intestinal tissue comparing Crohn's disease (CD), ulcerative colitis (UC) and control (C) patients. Subgroup analysis in ileal (CD n=17, UC n=5, C n=9) or colonic biopsies only (CD n=7, UC n=2, C n=7). D) OCTN2 protein expression in human intestinal tissue comparing CD, UC and C patients. Subgroup analysis in ileal (CD n=19, UC n=6, C n=10) or colonic (CD n=7, UC n=2, C n=11) biopsies only. Journal of Crohn's and Colitis 2012 6, 189-197DOI: (10.1016/j.crohns.2011.08.003) Copyright © 2011 European Crohn's and Colitis Organisation Terms and Conditions

Figure 3 Intestinal OCTN protein expression as a function of OCTN genotypes. Western blot analysis used tissue actin levels as a standard. A) OCTN1 protein expression in human intestinal tissue comparing control (C) and Crohn's disease (CD) patients with SLC22A4 CC, TC or TT genotypes. Expression of OCTN1 protein with CC (n=5) versus TT (n=8) genotypes (#p<0.05). B) OCTN2 protein expression in human intestinal tissue comparing C and CD patients with SLC22A5 GG, GC or CC genotypes. Expression of OCTN2 protein with GG (n=6) or CG (n=4) versus CC (n=4) genotypes (p<0.1). C) OCTN1 protein expression in human intestinal tissue comparing C and CD patients with wild type (CC) or mutated (TC-TT) (n=13) SLC22A4 genotypes (*p<0.025). D) OCTN2 protein expression in human intestinal tissue comparing C and CD patients with wild type (GG) or mutated (CG-CC) (n=8) SLC22A5 genotypes. E) OCTN1 protein expression in human intestinal tissue comparing all patients with SLC22A4 CC, TC or TT polymorphisms. Expression of OCTN1 protein with CC (n=14) versus TT (n=7) genotypes (and p<0.05). F) OCTN2 protein expression in human intestinal tissue comparing all patients with SLC22A5 GG (n=11), GC (n=8) or CC (n=8) genotypes. Journal of Crohn's and Colitis 2012 6, 189-197DOI: (10.1016/j.crohns.2011.08.003) Copyright © 2011 European Crohn's and Colitis Organisation Terms and Conditions

Figure 4 A) Carnitine transport in human intestinal brush border membrane vesicles. Results are presented according to the presence or absence of Na+. Counts per minute (CPM), Time (T) in seconds. B) Intestinal carnitine transport in Crohn's disease (CD) and control (C) patients. Carnitine transport was measured in human intestinal brush border membrane vesicles obtained from ileal resections in patients with CD (n=8) and C patients (n=9). Journal of Crohn's and Colitis 2012 6, 189-197DOI: (10.1016/j.crohns.2011.08.003) Copyright © 2011 European Crohn's and Colitis Organisation Terms and Conditions

Figure 5 Intestinal brush border membrane carnitine transport in Crohn's disease (CD) and control (C) patients comparing SLC22A4 (C-T substitution) and SLC22A5 (G-C transversion) polymorphisms. A) Comparison of intestinal carnitine transport between OCTN1 CC polymorphism group and TC and TT polymorphism groups in CD or C patients. B) Comparison of intestinal carnitine transport between OCTN1 CC polymorphism group and TC/TT polymorphism pooled group in CD or C patients. C) Comparison of intestinal carnitine transport between OCTN2 GG polymorphism and CG polymorphism groups in CD or C patients. C patient subgroup: GG (n=3) vs CG (n=2) (# p=0.001). Journal of Crohn's and Colitis 2012 6, 189-197DOI: (10.1016/j.crohns.2011.08.003) Copyright © 2011 European Crohn's and Colitis Organisation Terms and Conditions