1. Laser Capture Microdissection Reveals Transcriptional Abnormalities in Alopecia Areata before, during, and after Active Hair Loss 
Jane Li, Catherine van Vliet, Nicholas W. Rufaut, Leslie N. Jones, Rodney D. Sinclair, Francis R. Carbone 
Journal of Investigative Dermatology 
Volume 136, Issue 3, Pages (March 2016)
DOI: /j.jid
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2. Figure 1
Laser capture microdissection of hair follicles reveals abnormal gene expression in alopecia areata (AA). Healthy and AA bulbs and infundibulums were microdissected and mRNA extracted for PCR. Data obtained were normalized to housekeeping gene expression. (a) Representative active AA bulb laser capture microdissection. Scale bars = 75 μm. (b) Heatmap of significantly differentially expressed genes (DEGs) in bulbs and infundibulums after pairwise comparison of normal and active, regrown and unaffected AA groups (n = 24–44 bulbs/infundibulums). DEGs were determined by the Mann-Whitney U-test with multiple testing correction using the Benjamini-Hochberg method (P < 0.05). A red-green color scale shows relative gene expression (green = low, red = high). Heatmap generated using GENE-E software ( (c) Chemokine expression profile in normal hair follicles. Data presented as mean ± SEM. (d) Normalized gene expression levels of selected chemokine-chemokine receptor pairs in active AA bulbs. Spearman's r as shown (n = 37; all P < 0.01). (e) Venn diagram showing overlap of DEGs between normal bulbs and bulbs from active, regrown, and unaffected AA (n = 34–44 bulbs).
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions

3. Figure 2
Immunofluorescent staining of CD103, CST6, and ULBP3 in alopecia areata (AA) hair bulbs. Immunostaining images shown are representative of at least three independent experiments. Hoechst stain (blue) marks nuclei. Scale bars = 100 μm. (a) Expression of selected genes (green) upregulated in AA versus normal bulbs. (b) Immunostaining for CD3 (gray), CD8 (green), and CD103 (red), with the closeup view showing colocalization of CD8 and CD103. (c) ITGAE mRNA expression of bulbs from sequential scalp biopsies obtained during initial onset of AA. Biopsies collected from an active region and an unaffected region during periods of hair loss (t = 0–2 weeks; n = 7 active, 10 unaffected bulbs) and regrowth (t = 3 months; n = 6 regrown, 4 unaffected bulbs). ITGAE detected in three of six bulbs from the originally active area at later time point. (d) CST6 (red) immunostaining in normal and unaffected AA bulbs. (e) CST6 mRNA expression in active bulbs plotted by AA subtype (AT/AU = alopecia totalis/universalis; horizontal line indicates mean). (f) ULBP3 expression in normal and active AA bulbs by PCR (n indicates number of follicles; error bars show mean ± SEM) and (g) by immuno-staining (red; results representative of eight patients with AA and four normal controls).
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions