Association of ORAI1 gene polymorphisms with chronic spontaneous urticaria and the efficacy of the nonsedating H1 antihistamine desloratadine  Jie Li, MD, PhD, Aiyuan Guo, MD, PhD, Wangqing Chen, PhD, Lianghua Bin, PhD, Yijing He, MD, PhD, Wu Zhu, MD, PhD, Cong Peng, PhD, Siyu Yan, MD, Mingliang Chen, MD, PhD, Jianglin Zhang, MD, PhD, Juan Su, MD, PhD, Mei Yi, MD, PhD, Zhaoqian Liu, MD, PhD, Wei Zhang, MD, PhD, Weiqi Zeng, PhD, Donald Y.M. Leung, MD, PhD, Xiang Chen, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 139, Issue 4, Pages 1386-1388.e9 (April 2017) DOI: 10.1016/j.jaci.2016.10.017 Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 CSU susceptibility-associated ORAI1 SNPs have increased promoter activation and mast cell degranulation. A, ORAI1 promotor with rs12320939T allele has increased promoter activation than ORAI1 with rs12320939G allele determined by reporter gene assays. B, ORAI1 gene expression in PBMCs was evaluated by real-time PCR. Subjects who carried the rs12320939TT genotype (n = 7) have increased ORAI mRNA levels as compared with subjects who carried the rs12320939GG genotype (n = 5). C, The resting [Ca2+] influx (black bar) and the thapsigargin (TG) evoking [Ca2+] influx (gray bar) are significantly increased in RBL-2H3 cells that overexpressed the rs3741596A allele than RBL-2H3 cells that overexpressed the rs3741596G allele. D, The β-HEX release by phorbol 12-myristate 13-acetate and ionomycin stimulation was evaluated in RBL-2H3 cells with different rs3741596 alleles. Cells were first silenced for ORAI1 gene (siRNA-ORAI1) and then reconstituted with different rs3741596 alleles. Cells with rs3741596A (ORAI1-A, 0.81 ± 0.05) have significantly greater β-HEX release than do cells with rs3741596G (ORAI1-G, 0.61 ± 0.04). Data were presented as mean ± SD of at least 4 independent experiments performed in triplicate. β-HEX, β-Hexosaminidase. Journal of Allergy and Clinical Immunology 2017 139, 1386-1388.e9DOI: (10.1016/j.jaci.2016.10.017) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Site of each SNP in ORAI1 gene. Both rs12313273 and rs12320939 are located in the promoter region of ORAI1. rs3741595 and rs3741596 are located in the exon; the former results in a synonymous mutation, whereas the latter is a nonsynonymous mutation leading to the replacement of Serine by Glycine at amino acid 218. TSS, Transcription start site. Journal of Allergy and Clinical Immunology 2017 139, 1386-1388.e9DOI: (10.1016/j.jaci.2016.10.017) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 LD (linkage disequilibrium) heatmap of the linkage disequilibrium (D′, r2) among the 4 SNPs. Linkage disequilibrium was assessed through the Web site http://analysis.bio-x.cn/myAnalysis.php. r2 more than 0.8 means SNPs linkage disequilibrium. All r2 values were less than 0.8 between the different SNPs. Journal of Allergy and Clinical Immunology 2017 139, 1386-1388.e9DOI: (10.1016/j.jaci.2016.10.017) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions