A Promoter Sequence Variant of ZNF750 Is Linked with Familial Psoriasis Chi-Fan Yang, Wuh-Liang Hwu, Li-Cheng Yang, Wen-Hung Chung, Yin-Hsiu Chien, Chia-Fu.

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A Promoter Sequence Variant of ZNF750 Is Linked with Familial Psoriasis Chi-Fan Yang, Wuh-Liang Hwu, Li-Cheng Yang, Wen-Hung Chung, Yin-Hsiu Chien, Chia-Fu Hung, Hung-Chih Chen, Pei-Joung Tsai, Cathy S.J. Fann, Fang Liao, Yuan-Tsong Chen Journal of Investigative Dermatology Volume 128, Issue 7, Pages 1662-1668 (July 2008) DOI: 10.1038/jid.2008.1 Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Schematic representation showing four variants cosegregated with the disease in the family. The position and size of the genes are indicated by red squares. Nucleotide sequence and amino-acid translation of the patients show heterozygous variation marked by arrows. Journal of Investigative Dermatology 2008 128, 1662-1668DOI: (10.1038/jid.2008.1) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 The effect of p.Ala201Val variant in CD7. (a) Flow cytometric analysis of CD7 expression in gated CD4+ T cells, CD8+ T cells, and CD56+ NK cells from a representative normal control (upper panel) and his affected brother (lower panel). Histograms show surface fluorescence intensity of isotype-matched control (empty histogram) and anti-CD7 mAb (filled histogram). The percentage of CD7 expression was indicated. (b, c) PBMC cells from two patients (P1, P2) and five normal controls (N1–N5) were cultured with anti-CD3 or anti-CD3 plus anti-CD7 in a total volume of 200μl. The 2-day culture supernatants were collected and analyzed for production of (b) TNF-α and (c) IFN-γ by ELISA. (d) PBMC cells from two patients and five normal controls were cultured with anti-CD3 or anti-CD3 plus anti-CD7, and cell proliferation was performed by 3H-thymidine uptake. Journal of Investigative Dermatology 2008 128, 1662-1668DOI: (10.1038/jid.2008.1) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 The effect of the variations in the promoter region of ZNF750. The luciferase activity of the reporter constructs containing either the A or the C allele at the c.-625 locus in HaCaT cells. The results represent means±SD of five independent experiments. *Student's t-test. Journal of Investigative Dermatology 2008 128, 1662-1668DOI: (10.1038/jid.2008.1) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 EMSA showing binding of transcription factors to the -625C allele. (a) EMSAs were performed with HaCaT nuclear extract and the biotin-labeled 30-bp oligonucleotides containing the normal allele A or mutant allele C (lanes 1–4). Competition experiments were performed by using biotin-labeled oligonucleotides with mutant alleles and a 5-fold (lanes 5 and 8), 50-fold (lanes 6 and 9), and 100-fold (lanes 7 and 10) excess of unlabeled mutant allele (lanes 5–7) or normal allele (lanes 8–10). Arrows indicate the DNA–protein complex. (b) Competition assays. Lane 1 contained biotin-labeled mutant oligonucleotides with nuclear extract. Lanes 2–8 contained oligonucleotides with consensus binding sites for different transcription factors with fold concentrations as indicated. Journal of Investigative Dermatology 2008 128, 1662-1668DOI: (10.1038/jid.2008.1) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions