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Epistatic interaction between Toll-like receptor 3 (TLR3) and prostaglandin E receptor 3 (PTGER3) genes  Mayumi Ueta, MD, PhD, Gen Tamiya, PhD, Katsushi.

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Presentation on theme: "Epistatic interaction between Toll-like receptor 3 (TLR3) and prostaglandin E receptor 3 (PTGER3) genes  Mayumi Ueta, MD, PhD, Gen Tamiya, PhD, Katsushi."— Presentation transcript:

1 Epistatic interaction between Toll-like receptor 3 (TLR3) and prostaglandin E receptor 3 (PTGER3) genes  Mayumi Ueta, MD, PhD, Gen Tamiya, PhD, Katsushi Tokunaga, PhD, Chie Sotozono, MD, PhD, Masao Ueki, PhD, Hiromi Sawai, PhD, Tsutomu Inatomi, MD, PhD, Toshiyuki Matsuoka, MD, PhD, Shizuo Akira, MD, PhD, Shuh Narumiya, MD, PhD, Kei Tashiro, MD, PhD, Shigeru Kinoshita, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 129, Issue 5, Pages e11 (May 2012) DOI: /j.jaci Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Functional interaction between EP3 and TLR3. In TLR3/PTGER3 DKO mice the number of eosinophils in the lamina propria mucosae of the conjunctiva was decreased to a level similar to that seen in TLR3 KO mice and was significantly lower than that seen in either PTGER3 KO or wild-type mice. Data are shown as means ± SEMs of samples from all the mice examined (wild-type: phosphate-buffered saline, n = 24; RW, n = 28; PTGER3 KO mice: phosphate-buffered saline, n = 23; RW, n = 25; EP3/TLR3 DKO mice: phosphate-buffered saline, n = 4; RW, n = 11; TLR3 KO mice: phosphate-buffered saline, n = 12; RW, n = 12). ∗P < Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 LD in EP3 and TLR3 regions. LD in the TLR3 and PTGER3 regions show 3 solid-spine LD blocks in each region. Iterative SIS reported 14 variables with nonzero regression coefficients, as if connecting the 5′ region of TLR3 and the 3′ region of PTGER3. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Features of patients with SJS/TEN with ocular complications. A, Vesiculobullous lesions of the skin in the acute stage. B, Severe conjunctivitis in the acute stage. C, Oral involvement, including blisters, erosions, and bleeding of the mouth and lips in the acute stage. D, Paronychia in the acute stage. E, Conjunctival invasion into the cornea in the chronic stage. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Expression of TLR3 mRNA in conjunctival tissues from patients with SJS/TEN and chemical eye burn and the control subjects. Total RNA was isolated from conjunctival tissue sections by using the RNeasy mini kit (Qiagen, Hilden, Germany), according to the manufacturer’s instructions. The RT reaction was with the SuperScript preamplification kit (Invitrogen, Carlsbad, Calif). Quantitative RT-PCR was on an ABI-prism 7700 instrument (Applied Biosystems). The probes for human TLR3 and human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Applied Biosystems. The results were analyzed with sequence detection software (Applied Biosystems). The quantification data were normalized to the expression of the housekeeping gene GAPDH. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Immunohistologic analysis of TLR3 in conjunctival epithelium of patients with SJS/TEN and chemical eye burn and control subjects. For TLR3 staining, we used rabbit polyclonal antibody to TLR3 (Abcam, Cambridge, Mass). The secondary antibody (Biotin-SP–conjugated AffiniPure F[ab′]2 fragment donkey anti-rabbit IgG [H+L], 1:500 dilution; Jackson ImmunoResearch, Baltimore, Md) was applied for 30 minutes. The VECTASTAIN ABC reagent (Vector Laboratories, Burlingame, Calif) was used for increased sensitivity with peroxidase substrate solution (DAB substrate kit; Vector Laboratories) as a chromogenic substrate. Each scale bar represents 100 μm. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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