Hanna Niehues, Lam C. Tsoi, Danique A. van der Krieken, Patrick A. M

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Psoriasis-Associated Late Cornified Envelope (LCE) Proteins Have Antibacterial Activity Hanna Niehues, Lam C. Tsoi, Danique A. van der Krieken, Patrick A.M. Jansen, Merel A.W. Oortveld, Diana Rodijk-Olthuis, Ivonne M.J.J. van Vlijmen, Wiljan J.A.J. Hendriks, Richard W. Helder, Joke A. Bouwstra, Ellen H. van den Bogaard, Philip E. Stuart, Rajan P. Nair, James T. Elder, Patrick L.J.M. Zeeuwen, Joost Schalkwijk Journal of Investigative Dermatology Volume 137, Issue 11, Pages 2380-2388 (November 2017) DOI: 10.1016/j.jid.2017.06.003 Copyright © 2017 The Authors Terms and Conditions

Figure 1 In vivo eQTL analysis. (a) Quantile-quantile plot of rs4112788 versus expression traits in the epidermal differentiation complex region. The shaded area indicates 95% confidence interval of statistical significance. (b) Estimated effect size of LCE3B/C-del on in vivo LCE3A expression in the eQTL data set. Error bars = standard error of the mean. eQTL, expression quantitative trait loci; LCE, late cornified envelope; n.e., no expression; NN, normal skin; PN; nonlesional psoriatic skin; PP, lesional psoriatic skin. Journal of Investigative Dermatology 2017 137, 2380-2388DOI: (10.1016/j.jid.2017.06.003) Copyright © 2017 The Authors Terms and Conditions

Figure 2 In vitro mRNA analysis. mRNA expression of (a) control, (b) Th1-, and (c) Th17-cytokine stimulated cells (N = 5 per group) in wt/wt and del/del 3D reconstructed epidermis. (d, e) mRNA expression on Th1- and Th17-cytokine stimulation (N = 10). Error bars = standard error of the mean. Red lines = control (wt/wt or unstimulated, set to 1), *P < 0.05, **P < 0.005, ***P < 0.0005. LCE, late cornified envelope; Th1, T helper type 1; Th17, T helper type 17; wt, wild type. Journal of Investigative Dermatology 2017 137, 2380-2388DOI: (10.1016/j.jid.2017.06.003) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Barrier function of LCE3B/C wt/wt and del/del 3D reconstructed epidermis. (a–d) TEWL of unstimulated, Th1-, or Th17-stimulated epidermis. Repeated measures ANOVA plus Bonferroni post hoc with P < 0.05 significance. (e) Stratum corneum hydrocortisone-diffusion assay. (f) Lucifer yellow (LY) penetration in unstimulated, Th1-, or Th17-cytokine stimulated epidermis. Scale bar = 100 μm. N = 5 per genotype group. Error bars = standard error of the mean. ANOVA, analysis of variance; LCE, late cornified envelope; TEWL, transepidermal water loss; Th1, T helper type 1; Th17, T helper type 17; wt, wild type. Journal of Investigative Dermatology 2017 137, 2380-2388DOI: (10.1016/j.jid.2017.06.003) Copyright © 2017 The Authors Terms and Conditions

Figure 4 Antimicrobial activity of LCE3 proteins. LCE3A, B, C were incubated with Gram-negative (a–d) and Gram-positive commensal and pathogenic bacteria (e–j) between 0.1 to 10 μM protein. For specifications see Supplementary Table S4. 2 × 102 CFU/ml is the detection limit; therefore values <2 × 102 CFU/ml are not depicted. Error bars = standard error of the mean. CFU, colony forming units; LCE, late cornified envelope. Journal of Investigative Dermatology 2017 137, 2380-2388DOI: (10.1016/j.jid.2017.06.003) Copyright © 2017 The Authors Terms and Conditions