1. Receptor-interacting protein kinase 3 controls keratinocyte activation in a necroptosis- independent manner and promotes psoriatic dermatitis in mice 
Tetsuya Honda, MD, PhD, Osamu Yamamoto, MD, PhD, Yu Sawada, MD, PhD, Gyohei Egawa, MD, PhD, Akihiko Kitoh, MD, PhD, Atsushi Otsuka, MD, PhD, Teruki Dainichi, MD, PhD, Saeko Nakajima, MD, PhD, Yoshiki Miyachi, MD, PhD, Kenji Kabashima, MD, PhD 
Journal of Allergy and Clinical Immunology 
Volume 140, Issue 2, Pages e6 (August 2017)
DOI: /j.jaci
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2. Fig 1
RIPK3 expression in the epidermis of psoriatic lesions. A, Representative RIPK3 staining (red) of the psoriatic lesions (n = 20). Upper and lower right panels indicate higher magnification in the square of nonlesional and lesional areas of the left panel. Scale bars = 500 μm (left panel) and 200 μm (right panel). B, Ripk3 mRNA expression in the mouse ear skin with or without Aldara treatment for 3 days (n = 5). C, RIPK3 staining (brown) of mouse ear skin with or without Aldara treatment for 3 days (n = 10). Scale bars = 20 μm. D, An LDH assay using primary mouse keratinocytes. Keratinocytes were treated with Aldara at the indicated dilution for 8 hours, and the culture supernatants were subjected to an LDH assay (n = 3). E, TUNEL staining (red) of murine ear skin of Ripk3+/+ mice and Ripk3−/− mice treated with or without Aldara for 3 days. Blue represents nuclei stained by 4′-6-diamidino-2-phenylindole, dihydrochloride. Scale bars = 20 μm. Data are representative of at least 3 independent experiments and are expressed as mean ± SEM. *P < .05.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3. Fig 2
Impaired cytokine and chemokine expression from Ripk3−/− keratinocytes. A, The mRNA expression of CXCL2, IL-1β, and IL-24 in primary cultured keratinocytes from Ripk3+/+ or Ripk3−/− mice treated with Aldara for 8 hours (n = 3). B, The mRNA expression of CXCL2, IL-1β, and IL-24 in the ear skin treated with or without Aldara for 3 days (n = 4-6). C, Ear swelling response after Aldara treatment in Ripk3+/+ or Ripk3−/− mice (n = 4). D, Hematoxylin and eosin (HE)–stained skin section on day 3 of Aldara-treated or nontreated Ripk3+/+ mice and Ripk3−/− mice (n = 4). Scale bars = 50 μm. E and F, Flow cytometry analysis for measuring the number of neutrophils in the ear skin of Ripk3+/+ or Ripk3−/− mice (Fig 2, E) and bone marrow chimeric mice (Fig 2, F) treated with Aldara for 3 days (n = 4-6). Data are representative of at least 3 or 2 (Fig 2, F) independent experiments and are expressed as mean ± SEM. *P < .05.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4. Fig E1
A, Representative TUNEL staining (red) of murine ear skin of Ripk3+/+ mice treated with or without Aldara for 3 days. White dotted lines indicate the boundary between the epidermis and dermis. Scale bars = 20 μm. B, A representative picture of electron microscopic analysis of dead keratinocytes in the psoriasis skin lesion. Scale bar = 2 μm. Data are a representative of 8 (Fig E1, A) and 10 (Fig E1, B) independent samples.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5. Fig E2
Quantification of TUNEL-positive cells in epidermis of Ripk3+/+ mice and Ripk3−/− mice with or without Aldara treatment for 3 days (n = 4). Data are representative of 2 independent experiments with reproducible results and are expressed as mean ± SEM.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6. Fig E3
The mRNA expression of CXCL2, IL-1β, and IL-24 in epidermal sheet from Ripk3+/+ and Ripk3−/− mice treated with or without Aldara for 3 days (n = 3-5). Data are representative of 2 independent experiments with reproducible results and are expressed as mean ± SEM. *P < .05.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7. Fig E4
The mRNA expression of IL-17A, IL-23p19, and IL-22 in the ear skin of Ripk3+/+ mice and Ripk3−/− mice treated with or without Aldara for 3 days (n = 4-6). Data are representative of at least 3 independent experiments and are expressed as mean ± SEM.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8. Fig E5
The mRNA expression of CXCL2, IL-1β, and IL-24 in the ear skin of bone marrow chimeric mice treated with Aldara for 3 days (n = 4-6). Data are representative of 2 independent experiments with reproducible results and are expressed as mean ± SEM. *P < .05.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci )
Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions