Supplementary Figure 1 IMQ-Induced Mouse Model of Psoriasis. IMQ cream was
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1 Supplementary Figure 1 IMQ-Induced Mouse Model of Psoriasis. IMQ cream was painted on the shaved back skin of CBL/J and BALB/c mice for consecutive days. (a, b) Phenotypic presentation of mouse back skin treated with IMQ (severe erythema, scales and crusts). (c) H&E staining of lesional skin from treated mice. a, acanthosis; b, 1
2 increased proliferative basal layer epidermal keratinocytes; c, dilated capillaries (small blood vessels in dermis); d, dermal cell infiltrates; k, hyperkeratosis; m, microabscesses; r, elongated rete ridge. Dotted line indicates the border between the epidermis and dermis; scale bar, 100μm.
3 Supplementary Figure Decreased IL- levels in IL-1A -/- mice and IL--mediated activation of NF-κB Signaling in Keratinocytes. (a) qpcr analysis of IL- expression from skin of WT (n=) and IL-1A -/- mice (n=) treated with IMQ. Results are presented as the ratio of mrna to -actin, relative to that in IL-1A -/- mice. (b) mir-1 expression in HaCaT cells stimulated with IL- in absence or presence of various concentrations of STAT inhibitor (istat) (SI-01, Selleck #S11). Results are presented as the ratio of mirna to the small nuclear RNA U, relative to that in untreated cells. (c) Phosphorylated-p (P-p) and CD flow cytometry analysis of single cell suspensions from normal epidermis of mouse treated with vehicle (Ctr) or lesional epidermis of mouse treated with IMQ. **p 0.01, two-tailed Student s t-test. Error bars depict SEM.
4 9 Supplementary Figure Generation of mir-1 TG Mice. (a) Schematic illustration of the mir-1 transgenic construct used in this study to overexpress mir-1. CMV, cytomegalovirus (promoter); EGFP, enhanced green fluorescent protein. (b) PCR genotyping of mir-1 transgenic founder mice (bp). # indicated genotyping positive founder lines; m, DNA size marker; WT, wild-type control. (c) Quantitative RT-PCR analysis of mir-1 in PBMCs derived from WT or mir-1 TG mice (n= for each group). Results are presented as the ratio of mirna to the small nuclear RNA U, relative to that in WT mice. *p 0.0, two-tailed Student s t-test. Error bars depict SEM.
5 Supplementary Figure Increased Disease Severity in mir-1 TG Mice Treated with IMQ. (a, b) H&E staining of the back skin of WT or mir-1 TG mice treated with IMQ. (c) Digital photos of H&E-stained skin samples derived from WT or mir-1 TG mice treated with IMQ were taken at the same orientation and magnification (n=). The epidermal area was outlined, and its pixel area was measured using the lasso tool in Adobe Photoshop CS. The relative area of the epidermis was calculated by the formula provided in Methods. Scale bar, 100μm. (d) Dermal cell infiltrates for WT or mir-1 TG mice treated with IMQ. For all measurements, the median number of specifically stained dermal nucleated cells was counted in high-power fields per section. *p 0.0, **p<0.01, two-tailed Student s t-test. Error bars depict SEM. 1
6 Supplementary Figure Enhanced Proliferation of NHEK after overexpressing mir-1. Cell cycle analysis of NHEK transfected with control mimics (Ctr) and mir-1 mimics. Data are representative of two independent experiments.
7 Supplementary Figure Decreased Disease Severity in cko mice after IL- Treatment. Mouse recombinant IL- was injected intradermally into the ear skin of mir-1 fl/fl (n=) and cko mice (n=) every other day for 1 days. (a) Phenotype of ear skin from mir-1 fl/fl and cko mice treated with ril-. (b) H&E staining of lesional skin from mir-1 fl/fl and cko mice treated with ril-. Scale bar, 100μm. (c, d) Skin thickness and acanthosis were quantitated from the H&E staining. *p 0.0, two-tailed Student s t-test. Error bars depict SEM.
8 Supplementary Figure Expression Levels of Inflammatory Genes in Lesional Skin of IMQ-Induced and IL--Mediated Mouse Models of Psoriasis. (a-f) Expression levels of IL-1, IL-, IL-1A, IL-, IFN- and TNF- in lesional skin samples derived from either mir-1 fl/fl or cko mice treated with IMQ. (g-l) Expression levels of IL-1,
9 IL-, IL-1, IL-, IFN- and TNF- in lesional ear skin samples derived from either mir-1 fl/fl or cko mice treated with ril-. Data are representative of two independent experiments. ns, not significant, *p 0.0, two-tailed Student s t-test. Error bars depict SEM. 9
10 Supplementary Figure mir-1 Expression in Epidermal Cells and Splenocytes of mir-1 fl/fl and cko Mice Treated with IMQ. (a) Expression of mir-1 in epidermis from mir-1 fl/fl and cko mice treated with vehicle (Ctr) or IMQ (n= for each group). (b) Expression of mir-1 in splenocytes from mir-1 fl/fl and cko mice treated with vehicle (Ctr) or IMQ (n= for each group). Results are presented as the ratio of mirna to the small nuclear RNA U, relative to that in mir-1 fl/fl mice treated with vehicle. ns, not significant, *p 0.0, two-tailed Student s t-test. Error bars depict SEM. 9 10
11 Supplementary Figure 9 Identification of mir-1 Target Gene(s). (a, b) Predicted 0 candidate targets of mir-1. RNA was extracted from skin samples of non-treated WT mice (Control), WT mice treated with IMQ (WT) and mir-1 TG mice treated with IMQ (mir-1 TG ). (c) mrna levels of 1 candidate genes were measured by qpcr, and the 11
12 rest genes were undetectable. Results are presented as the ratio of mrna to -actin, relative to that in untreated WT controls. *p 0.0, **p 0.01, ***p<0.001, two-tailed Student s t-test. Error bars depict SEM. 1
13 Supplementary Figure 10 Upregulated Pppc Levels in cko Mice in IL--mediated Mouse Model of Psoriasis. (a) qpcr analysis of pppc expression in ear skin of mir-1 fl/fl and cko mice, treated with ril- or PBS (n=-). Results are presented as the ratio of mrna to -actin, relative to that in mir-1 fl/fl mice treated with PBS. (b) Western blotting of pppc expression in ear skin of mir-1 fl/fl and cko mice, treated with ril- or PBS. Data are representative of two independent experiments. **p 0.01, two-tailed Student s t-test. Error bars depict SEM. 1
14 9 Supplementary Figure 11 Pppc is not Targeted by mir-1a and Silencing Pppc does not alter mir-1 expression. (a, b) Ago was immunoprecipitated from epidermis lysates derived from mir-1 fl/fl or cko mice treated with IMQ. Immunoprecipitates were assayed for pppc and mir-1a. (c) Expressin of mir-1 in NHEK transfected with non-targeted sirna (sirna-ctr) and pppc targeted sirna (sirna-pppc). Results are presented as the ratio of mrna to -actin, relative to that in NHEK transfected with non-targeted sirna. Data are representative of two independent experiments. ns, not significant, two-tailed Student s t-test. Error bars depict SEM. 10 1
15 9 Supplementary Figure 1 IL-1A Inhibits Pppc Expression in Keratinocytes. (a) Western blotting of pppc expression in primary mouse keratinocytes stimulated by IL-1A with different dosages and time points. (b, c) Western blotting of pppc expression in epidermis derived from IL-1A +/+ or IL-1A -/- mice treated with IMQ for days. Values (a, b) were expressed as fold changes relative to non-stimulated keratinocytes (a) or to IL-1A +/+ mouse (b). *p 0.0, two-tailed Student s t-test. Error bars depict SEM. 10 1
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17 1
18 Supplementary Figure 1 List of original pictures of western blots. Black boxes highlight the indicated lanes in figures. 1
19 Supplementary Table 1. Information for patients with psoriasis vulgaris Sample ID Age in Gender PASI Score Years 1 1 M. M 1. M 1. M 10. M 1 19 F 1. F 1. 1 M unknown 9 M F. 11 M 9. 1 M unknown 1 M M M unknown 1 F M 1 1 M. 19 F M 1. 1 M 10. M 1. F 11. M. 9 M 1. M.9 M 1. 9 M. 9 M 1.9 All patients were clinically diagnosed as psoriasis vulgaris. M, male; F, female; PASI, Psoriasis Area and Severity Index; Unknown, data was missing. 19
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SUPPLEMENTARY FIGURES AND TABLE Supplementary Figure S1: Characterization of IRE1α mutants. A. U87-LUC cells were transduced with the lentiviral vector containing the GFP sequence (U87-LUC Tet-ON GFP).
well for 2 h at rt. Each dot represents an individual mouse and bar is the mean ±
Supplementary data: Control DC Blimp-1 ko DC 8 6 4 2-2 IL-1β p=.5 medium 8 6 4 2 IL-2 Medium p=.16 8 6 4 2 IL-6 medium p=.3 5 4 3 2 1-1 medium IL-1 n.s. 25 2 15 1 5 IL-12(p7) p=.15 5 IFNγ p=.65 4 3 2 1
Plasma exposure levels from individual mice 4 hours post IP administration at the
Supplemental Figure Legends Figure S1. Plasma exposure levels of MKC-3946 in mice. Plasma exposure levels from individual mice 4 hours post IP administration at the indicated dose mg/kg. Data represent
Title of file for HTML: Supplementary Information Description: Supplementary Figures and Supplementary Table
Title of file for HTML: Supplementary Information Description: Supplementary Figures and Supplementary Table Title of file for HTML: Peer Review File Description: Innate Scavenger Receptor-A regulates
Supplementary Figure 1
Supplementary Figure 1 YAP negatively regulates IFN- signaling. (a) Immunoblot analysis of Yap knockdown efficiency with sh-yap (#1 to #4 independent constructs) in Raw264.7 cells. (b) IFN- -Luc and PRDs
GFP/Iba1/GFAP. Brain. Liver. Kidney. Lung. Hoechst/Iba1/TLR9!
Supplementary information a +KA Relative expression d! Tlr9 5!! 5! NSC Neuron Astrocyte Microglia! 5! Tlr7!!!! NSC Neuron Astrocyte! GFP/Sβ/! Iba/Hoechst Microglia e Hoechst/Iba/TLR9! GFP/Iba/GFAP f Brain
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DOI: 1.138/ncb3355 a S1A8 + cells/ total.1.8.6.4.2 b S1A8/?-Actin c % T-cell proliferation 3 25 2 15 1 5 T cells Supplementary Figure 1 Inter-tumoral heterogeneity of MDSC accumulation in mammary tumor
DOI: 10.1038/ncb2210 b. ICAM1 ng ml -1 P = 0.0001 Small RNA (15-30nts) ng ml -1 Cell Lysate Exosome HDL Plasma HDL Normal Human HDL mirnas R = 0.45 P < 0.0001 Normal Human Exosome mirnas Figure S1. Characterization
Supplementary Figure 1
d f a IL7 b IL GATA RORγt h HDM IL IL7 PBS Ilra R7 PBS HDM Ilra R7 HDM Foxp Foxp Ilra R7 HDM HDM Ilra R7 HDM. 9..79. CD + FOXP + T reg cell CD + FOXP T conv cell PBS Ilra R7 PBS HDM Ilra R7 HDM CD + FOXP
T H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Krenn et al., http://www.jcb.org/cgi/content/full/jcb.201110013/dc1 Figure S1. Levels of expressed proteins and demonstration that C-terminal
Crucial role for human Toll-like receptor 4 in the development of contact allergy to nickel
Supplementary Figures 1-8 Crucial role for human Toll-like receptor 4 in the development of contact allergy to nickel Marc Schmidt 1,2, Badrinarayanan Raghavan 1,2, Verena Müller 1,2, Thomas Vogl 3, György
Nature Immunology: doi: /ni Supplementary Figure 1. Cytokine pattern in skin in response to urushiol.
Supplementary Figure 1 Cytokine pattern in skin in response to urushiol. Wild-type (WT) and CD1a-tg mice (n = 3 per group) were sensitized and challenged with urushiol (uru) or vehicle (veh). Quantitative
Kidney. Heart. Lung. Sirt1. Gapdh. Mouse IgG DAPI. Rabbit IgG DAPI
a e Na V 1.5 Ad-LacZ Ad- 110KD b Scn5a/ (relative to Ad-LacZ) f 150 100 50 0 p = 0.65 Ad-LacZ Ad- c Heart Lung Kidney Spleen 110KD d fl/fl c -/- DAPI 20 µm Na v 1.5 250KD fl/fl Rabbit IgG DAPI fl/fl Mouse
Integrin CD11b negatively regulates TLR-triggered inflammatory responses by. activating Syk and promoting MyD88 and TRIF degradation via cbl-b
Integrin CD11b negatively regulates TLR-triggered inflammatory responses by activating Syk and promoting MyD88 and TRIF degradation via cbl-b Chaofeng Han, Jing Jin, Sheng Xu, Haibo Liu, Nan Li, and Xuetao
Supplementary Figure 1 The ability to regenerate an ear hole is discontinuous with wound healing. Ear-hole closure at D85 for each sex within each
Supplementary Figure 1 The ability to regenerate an ear hole is discontinuous with wound healing. Ear-hole closure at D85 for each sex within each species observed. Data show a binary response to a 4 mm
Nature Immunology: doi: /ni Supplementary Figure 1. Production of cytokines and chemokines after vaginal HSV-2 infection.
Supplementary Figure 1 Production of cytokines and chemokines after vaginal HSV-2 infection. C57BL/6 mice were (a) treated intravaginally with 20 µl of PBS or infected with 6.7x10 4 pfu of HSV-2 in the
RAW264.7 cells stably expressing control shrna (Con) or GSK3b-specific shrna (sh-
1 a b Supplementary Figure 1. Effects of GSK3b knockdown on poly I:C-induced cytokine production. RAW264.7 cells stably expressing control shrna (Con) or GSK3b-specific shrna (sh- GSK3b) were stimulated
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SUPPLEMENTARY MATERIAL IL-1 signaling modulates activation of STAT transcription factors to antagonize retinoic acid signaling and control the T H 17 cell it reg cell balance Rajatava Basu 1,5, Sarah K.
Supplemental Materials for. Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to. FTY720 during neuroinflammation
Supplemental Materials for Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to FTY7 during neuroinflammation This file includes: Supplemental Table 1. EAE clinical parameters of
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DOI: 1.138/ncb222 / b. WB anti- WB anti- ulin Mitotic index (%) 14 1 6 2 T (h) 32 48-1 1 2 3 4 6-1 4 16 22 28 3 33 e. 6 4 2 Time (min) 1-6- 11-1 > 1 % cells Figure S1 depletion leads to mitotic defects
Supporting Information
Supporting Information Franco et al. 10.1073/pnas.1015557108 SI Materials and Methods Drug Administration. PD352901 was dissolved in 0.5% (wt/vol) hydroxyl-propyl-methylcellulose, 0.2% (vol/vol) Tween
SUPPLEMENTARY INFORMATION. Supp. Fig. 1. Autoimmunity. Tolerance APC APC. T cell. T cell. doi: /nature06253 ICOS ICOS TCR CD28 TCR CD28
Supp. Fig. 1 a APC b APC ICOS ICOS TCR CD28 mir P TCR CD28 P T cell Tolerance Roquin WT SG Icos mrna T cell Autoimmunity Roquin M199R SG Icos mrna www.nature.com/nature 1 Supp. Fig. 2 CD4 + CD44 low CD4
Control GST GST-RAP. α2-mg. 170 kda. b-actin. 42 kda LRP-1
% of max Supplementary Figure 1 Control GST GST-RP 17 kda α2-mg 42 kda b-actin Gate: CD11c+ (DCs) Gate: F4/8+ (Mfs) IgG Cd11cCre + Lrp1 fl/fl LRP-1 Supplementary figure 1. () MDCs were pretreated with
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DOI: 10.1038/ncb2566 Figure S1 CDKL5 protein expression pattern and localization in mouse brain. (a) Multiple-tissue western blot from a postnatal day (P) 21 mouse probed with an antibody against CDKL5.
Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the
Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the targeted allele in ES cells, and the mutant allele in
Supplementary Figure 1. Baf60c and baf180 are induced during cardiac regeneration in zebrafish. RNA in situ hybridization was performed on paraffin
Supplementary Figure 1. Baf60c and baf180 are induced during cardiac regeneration in zebrafish. RNA in situ hybridization was performed on paraffin sections from sham-operated adult hearts (a and i) and