1. Reversal of Murine Epidermal Atrophy by Topical Modulation of Calcium Signaling 
Basile Darbellay, Laurent Barnes, Wolf-Henning Boehncke, Jean-Hilaire Saurat, Gürkan Kaya 
Journal of Investigative Dermatology 
Volume 134, Issue 6, Pages (June 2014)
DOI: /jid
Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Store-Operated Calcium Entry (SOCE) is compromised in epidermal atrophy. Histological sections of healthy human skin (a) and atrophic skin (b) stained for Orai1 and Ki67; A=basal layer, B=suprabasal layers. (c) Quantification of Orai1 expression determined by the intensity of staining in a and b (mean±SD, see Materials and Methods). (d) Quantification of the Ki67-positive nuclei per 2mm of the dermo–epidermal junction (mean±SD, see Materials and Methods). (e) Cytoplasmic Ca2+ assessed by Fura-2 probe 24hours after isolation of primary keratinocytes. Intracellular Ca2+ stores were depleted using 2μM thapsigargin (Tg) in a medium containing 250nM Ca2+, and, subsequently, 1.8mM Ca2+ was added to the external medium to reveal SOCE (one representative experiment). (f) Peak Tg-induced SOCE amplitude measured in five patients/condition (mean±SD). Bar=150μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Orai1 inhibition results in epidermal atrophy in mice. (a) Histological sections of mouse skin stained for Orai1, hematoxylin–eosin (HE), and Ki67 (left panel, E=epidermis, D=dermis). Quantification of Orai1 staining intensity, epidermal thickness, and Ki67-positive nuclei standardized to control (right panel, mean±SD). Mice were treated with topical small interfering RNA (siRNA) (2 × per day, 2 weeks, see Materials and Methods). (b) Histological sections of mouse skin stained with HE and for Ki67 (left panel). Quantification of epidermal thickness standardized to control and Ki67-positive nuclei (right panel, mean±SD). Mice were treated with topical BTP2 (2 × per day, 2 weeks). (c) Ki67-positive nuclei per field at × 40 plotted with the level of expression of Orai1 (mean staining value per field at × 40) in mice treated with siRNA. Ki67 and Orai1 expressions are normalized. Bar=150μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
HB-EGF-induced proliferation relies on Orai1 in primary human keratinocytes (PHKs). Fura-2 fluorescence at 360-nm excitation measured in PHKs: (a) experiment was performed 2 days after transfection with siRNA; (b) BTP2 was applied 10minutes before the addition of Mn2+ (left panels: one representative experiment, right panels: normalized mean±SD in three independent experiments). (c) Proliferation of PHKs assessed by cell counting (see Methods). HB-EGF 5ngml−1 and BTP2 1μM were added in the culture medium. Day 0 is 48hours after small interfering RNA (siRNA) transfection (mean±SD in 10 fields/condition in three independent experiments).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Benzohydroquinone (BHQ)-induced Ca2+ influx through Orai1 elicits epidermal hyperplasia in mice and stimulates human keratinocyte proliferation. (a, b) Fura-2 fluorescence at 360-nm excitation measured in primary human keratinocytes (PHKs; as in Figure 3a and b). (c) Proliferation of PHKs assessed by cell counting (see Methods). HB-EGF 5ngml−1 and BTP2 1μM were added in the culture medium. BHQ 20μM was applied 2 × per day for 45minutes. Day 0 is 48hours after small interfering RNA (siRNA) transfection. (d) Fura-2 fluorescence at 360-nm excitation measured in primary murine keratinocytes. BTP2 was applied 10minutes before Mn2+ (as in Figure 3a and b). (e) Histological sections of mouse skin treated with solvent, BHQ±BTP2 (2 × per day, 2 weeks) stained with hematoxylin–eosin and for Ki67 (left panel). Quantification of epidermal thickness standardized to control and Ki67-positive nuclei (right panels, mean±SD on 10 fields/mouse). Bar=150μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Corticosteroid-induced skin atrophy is reversed by topical application of benzohydroquinone (BHQ). (a) Histological sections of mouse skin treated with solvent (control=ethanol), clobetasol 0.05% (2 × per day, 5 days), or clobetasol+BHQ 20μM (2 × per day, 5 days) stained with hematoxylin–eosin and for Ki67 (left panel). Mice were treated with clobetasol propionate (CP) for 5 days to induce epidermal atrophy and then for 5 additional days with CP+BHQ. (b) Quantification of epidermal thickness standardized to control and Ki67-positive nuclei (right panels, mean±SD, 10 fields/mouse). Bar=150μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2014 The Society for Investigative Dermatology, Inc Terms and Conditions