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. 2016 Aug 5;6:31119. doi: 10.1038/srep31119

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Copyright © 2016, The Author(s)

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(a) Normal human keratinocytes were obtained from adult foreskin, and peripheral blood neutrophils were isolated from 11 healthy controls, 7 eczema patients and 9 psoriasis patients. Normal human keratinocytes were treated with PMA or DNase I alone, or co-cultured for 16 hours with unstimulated control neutrophils (n = 11), control neutrophils stimulated with PMA (n = 11), control neutrophils treated with both PMA and DNase I (n = 11), unstimulated eczema neutrophils (n = 7), eczema neutrophils stimulated with PMA (n = 7), eczema neutrophils treated with both PMA and DNase I (n = 7), psoriatic neutrophils (n = 9), and psoriatic neutrophils treated with DNase I (n = 9). HBD-2 mRNA expression from keratinocytes was determined by real-time quantitative RT-PCR. ***P < 0.001, ANOVA with Bonferroni correction. (b) Netting neutrophils from psoriasis patients induced HBD-2 protein secretion by epidermal keratinocytes. Normal human keratinocytes were obtained from adult foreskin, and peripheral blood neutrophils were isolated from 7 healthy controls and 7 psoriasis patients. Normal human keratinocytes were treated with PMA or DNase I alone, or co-cultured for 24 hours with unstimulated control neutrophils (n = 7), control neutrophils stimulated with PMA (n = 7), control neutrophils treated with both PMA and DNase I (n = 7), psoriatic neutrophils (n = 7), and psoriatic neutrophils treated with DNase I (n = 7). HBD-2 protein secretion from keratinocytes was determined by ELISA. ***P < 0.001, ANOVA with Bonferroni correction.