Supplementary Figure 1. Antibiotic partially rescues mice from sepsis. (ab) BALB/c mice under CLP were treated with antibiotic or PBS.
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2 Supplementary Figure 1. Antibiotic partially rescues mice from sepsis. (ab) BALB/c mice under CLP were treated with antibiotic or PBS. (a) Survival curves. WT Sham (n=5), WT CLP or WT CLP antibiotic (n=20). (b) Bacterial load in the blood after CLP. (c-d) Serum and lungs of C57BL/6J mice under CLP were collected at the indicated times points. IL-1β, IL-6 and IL-10 concentrations in serum (c) and lung tissue (d) were determined by ELISA.(e) Serum endocan concentration was determined by ELISA and the levels of creatine kinase-mb (CK-MB), alanine aminotransferase (ALT) and blood urea nitrogen (BUN) were determined by colorimetric assays. ND, not detected. *p < 0.05 compare with naïve control or day 0 (b) (Mantel-Cox log-rank test in a, Mann-Whitney U test in b and one-way ANOVA result with Dunnett posthoc tests in c-e). Data are representative of three (a-b) or two (a, b, g) (mean ± s.e.m. in c-e and median in b). 2
3 Supplementary Figure 2. Kinetics of type-2 cytokines production during CLP. Lungs and bronchial lavage (BAL) fluids of mice under CLP and treated with antibiotic were collected at the indicated times points. (a) IL-4 and IL-13 concentrations in the lung tissues of C57BL/6J were determined by ELISA. (b) IL-33 concentrations in the BAL of BALB/c and Il1rl1 -/- at day 15 after CLP were determined by ELISA. *p < 0.05 and **p < 0.01, comparing groups as indicated or with day 0 (one-way ANOVA result with Dunnett posthoc tests in a and twotailed unpaired Student s t-test in b). Data are mean ± sem of 3-10 mice/group. 3
4 Supplementary Figure 3. Proinflammatory cytokines production during secondary infection in sepsis-surviving mice. C57BL/6J mice were subjected to CLP and challenged i.n. 15 days later with L. pneumophila. TNF and IL-6 concentrations in the BAL were determined by ELISA 12 h after challenge. ND, not detected. *p < 0.05 (two-tailed unpaired Student s t-test), ns, not significant. Data are mean ± sem of 3-5 mice/group. 4
5 Supplementary Figure 4. Repeated administration of IL-33 impaired bacterial clearance and increased type-2 cytokines production. BALB/c mice were inoculated i.n. with 1 or 3 µg of IL-33 or PBS for 4 consecutive days and challenged i.n. 2 days later with L. pneumophila. (a) Bacterial load in the lung tissue 48 h after challenge (n = 5 mice per group). (b) IL-4 and IL-13 concentrations in the lung tissue 48 h after challenge (n = 5 mice per group). *p < 0.05 (Mann-Whitney U test in a and two-tailed unpaired Student s t-test in b), ns, not significant. Data are median (a) and mean ± sem (b) of 5 mice/group. 5
6 Supplementary Figure 5. WT, Il1rl1 /, Stat6 -/- and Il10 -/- mice show similar survival curves following CLP and antibiotic treatment. Survival curves after CLP of (a) BALB/c and Il1rl1 / mice (n = 40 for WT group and n = 45 mice for Il1rl1 -/- CLP group), (b) BALB/c mice treated with sst2 or PBS (n = 15 mice per group), (c) BALB/c and Stat6 -/- mice (n = 40 for WT group and n = 42 mice for Stat6 -/- CLP group), and (d) C57BL/6J and Il10 -/- mice (n = 40 for WT group and n = 36 mice for Il10 -/- CLP group). Data are percentage of survival from 1 experiment (b) or pooled from three (d), five (c) or six (a) experiments. 6
7 Supplementary Figure 6. Induction of ILC2 in Rag1 / sepsis-surviving mice. Lungs of C57BL/6J and Rag1 -/- mice under CLP and antibiotic treatment were collected on day 3 after CLP. (a) Representative FACS plots of ILC2 + cells (Sca1 + in Lin - CD45 + cells) from BALB/c and C57BL/6J mice was determined by FACS (n 3 mice per group). Representative FACS plots from one experiment. (b) Representative FACS plots and frequency of ILC2 + cells (Sca1 + in Lin - CD45 + cells) from C57BL/6J and Rag1 -/- mice was determined by FACS (n 3 mice per group). *p < 0.05 (two-tailed unpaired Student s t-test in b), ns, not significant. Data are mean ± sem of 3-5 mice/group. 7
8 Supplementary Figure 7. M2 polarization with IL-4 IL-33 was abolished in Stat6 -/- BMDM in vitro. BMDM from BALB/c, BALB/c Stat6 -/- or BALB/c Il1rl1 -/- mice were cultured for 2 days in the presence of IL-4 and/or IL-33, or medium alone. (a) Percentage of F4/80 + CD206 + cells was analysed by FACS (each cell type was pooled from 2 mice and from 2 well per group). (b) Concentrations of CCL22 in the 2 days culture supernatants determined by ELISA (each cell type was pooled from 2 mice and 3-5 well per group). ND, not detected. *p < 0.05 (two-tailed unpaired Student s t-test in b), ns, not significant. 8
9 Supplementary Figure 8. Production of urea, IGF-1 and TGF-β of macrophages from sepsis-surviving mice. Peritoneal lavage fluid and lungs from BALB/c, Il1rl1 / and Stat6 -/- mice under CLP and antibiotic treatment were collected on day 15 after CLP. (a-b) Urea concentrations in the peritoneal lavage (a) and BAL (b) were determined by colorimetric assays. (c) IGF-1 concentrations in the lung tissue were determined by ELISA. *p < 0.05 and **p < 0.01 (two-tailed unpaired Student s t-test). Data are representative of two independent experiments (mean ± sem of 3 8 mice/group). 9
10 Supplementary Figure 9. M2 macrophage induces expansion of Treg cell population in sepsis-surviving mice. PBS or M2 macrophages were adoptively transferred (4x10 6 cells, i.v., on day 3 after CLP) into BALB/c or Stat6 -/- sepsis-surviving mice. The surviving mice were either sacrificed or challenged with L. pneumophila on day 15 after CLP. (a) Representative FACS plots, frequency and number of Foxp3 + CD4 + T cells (n = 3 mice per group). (b) Survival curves of mice after L. pneumophila challenge (n = 10 mice per group). *p < 0.05 and **p < 0.01 (two-tailed unpaired Student s t test in a, Mantel-Cox log-rank test in b). Data are representative of two (a) independent experiments or are pooled of two (b) experiments (mean ± s.e.m. in a). 10
11 Supplementary Figure 10. IL-10 production in the lungs of sepsissurviving mice. (a) Lung tissues from C57BL/6J mice under CLP and antibiotic treatment were collected at the indicated times points and IL-10 concentrations determined by ELISA.(b-d) Lung tissues of C57BL/6J, Rag1 -/- (b), BALB/c, Il1rl1 -/- and Stat6 -/- (c-d) mice under CLP and antibiotic treatment were collected on day 15 after CLP and IL-10 concentrations determined by ELISA. *p < 0.05, ***p < comparing groups as indicated or day 0 (a) (one-way ANOVA result with Dunnett posthoc tests in a and two-tailed unpaired Student s t-test in b-d). Data are representative of two independent experiments (mean ± sem of 3 10 mice/group). 11
12 Supplementary Figure 11. C57BL/6J and Il10 -/- mice produced similar levels of type-2 cytokines and frequency of M2 macrophages during sepsis. Lungs and peritoneal lavage fluids from C57BL/6J and Il10 / mice under CLP and antibiotic treatment were collected on day 15 after CLP. IL-4 (a) and IL-33 (b) concentrations in the lung tissue were determined by ELISA. (c) Frequency of peritoneal F4/80 + CD206 + macrophages was determined by FACS. *p < 0.05 (two-tailed unpaired Student s t-test), ns, not significant. Data are representative of two independent experiments (mean ± sem of 3 12 mice/group). 12
13 Supplementary Figure 12. Differential cell counts of the peripheral blood of septic patients. Peripheral blood from sepsis-surviving patients (n=11) and healthy controls (n = 14) was examined for (a) leukocytes, (b) granulocytes, (c) monocytes and (d) lymphocytes. Data are mean ± s.e.m. *p < 0.05 (two-tailed unpaired Student s t-test), ns, not significant. 13
14 Supplementary Figure 13. Full scan of the original uncropped western blot. Original Western blots used in Fig. 4e. (a) Western blot for Arg1. (b) Western blot for GAPDH. (c) Protein Ladder. n, naïve mice (n=9 mice). c, CLP mice (n=5 mice). 14
15 Supplementary Figure 14. Representative gating strategies used for flow cytometry analysis. Doublets were excluded by FSC-H and FSC-A gating for all flow cytometry analysis. (a) Proportion of ILC2 cells stained for ST2 + Lin - (CD3 - CD11c - CD19 - F4/80 - ) among CD45 + cells in single cell suspensions of lung cells. (b) Proportion of M2 macrophages stained for CD206 + and F4/80 + in peritoneal cells. (c) Proportion of Treg cells stained for FOXP3 + among CD4 + cells in spleen cells. 15
16 Supplementary Table 1 Demographic and clinical characteristics of septic patients Patients n=11 Age (years) - mean (SD) 60.2 (±12.83) Female/Male 4/7 APACHE II - mean (SD) (±9.73) SOFA - mean (SD) 9.81 (±4.87) SAP3 - mean (SD) 45.7 (±23.98) SD: standard deviation, APACHE II: Acute Physiology and Chronic Health Evaluation II, SOFA: Sepsis-related Organ Failure Assessment, SAP3: Simplified Acute Physiology score 3. 16
17 Supplementary Table 2 Clinical characteristics of septic patients Patients Site of Infection Time after sepsis (months) Severity of sepsis Microorganisms isolated 1 Respiratory 5 Septic Shock Staphylococcus haemolyticus 2 Respiratory 7 Severe Sepsis Enterococcus faecium 3 Respiratory 7 Severe Sepsis Escherichia coli 4 Respiratory 10 Septic Shock Streptococcus pneumoniae 5 Respiratory 9 Severe Sepsis Negative 6 Respiratory 7 Septic Shock Negative 7 Respiratory 6 Severe Sepsis Negative 8 Respiratory 5 Septic Shock Staphylococcus haemolyticus 9 Skin 5 Septic Shock Negative 10 Urinary 9 Severe Sepsis Staphylococcus aureus 11 Respiratory 7 Severe Sepsis Negative 17
Supplementary Figure 1. Characterization of basophils after reconstitution of SCID mice
Supplementary figure legends Supplementary Figure 1. Characterization of after reconstitution of SCID mice with CD4 + CD62L + T cells. (A-C) SCID mice (n = 6 / group) were reconstituted with 2 x 1 6 CD4
Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured
Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured under Th0, Th1, Th2, Th17, and Treg conditions. mrna
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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
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Supplemental Table 1: Demographics and characteristics of study participants. Male, n (%) 3 (20%) 6 (50%) Age, years [mean ± SD] 33.3 ± ± 9.
![Supplemental Table 1: Demographics and characteristics of study participants. Male, n (%) 3 (20%) 6 (50%) Age, years [mean ± SD] 33.3 ± ± 9.](/thumbs/90/103696086.jpg "Supplemental Table 1: Demographics and characteristics of study participants. Male, n (%) 3 (20%) 6 (50%) Age, years [mean ± SD] 33.3 ± ± 9.")
SUPPLEMENTAL DATA Supplemental Table 1: Demographics and characteristics of study participants Lean (n=15) Obese (n=12) Male, n (%) 3 (20%) 6 (50%) Age, years [mean ± SD] 33.3 ± 9.5 44.8 ± 9.1 White, n
Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and
Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and Thy1 in NH cells derived from the lungs of naïve mice.
Nature Medicine: doi: /nm.3922
Title: Glucocorticoid-induced tumor necrosis factor receptor-related protein co-stimulation facilitates tumor regression by inducing IL-9-producing helper T cells Authors: Il-Kyu Kim, Byung-Seok Kim, Choong-Hyun
Supplementary Figure 1. Efficiency of Mll4 deletion and its effect on T cell populations in the periphery. Nature Immunology: doi: /ni.
Supplementary Figure 1 Efficiency of Mll4 deletion and its effect on T cell populations in the periphery. Expression of Mll4 floxed alleles (16-19) in naive CD4 + T cells isolated from lymph nodes and
pro-b large pre-b small pre-b CCCP (µm) Rag1 -/- ;33.C9HCki
a TMRM FI (Median) b TMRM FI (Median) c 20 15 10 5 0 8 6 4 2 0 pro-b large pre-b small pre-b 0 10 20 30 40 50 60 70 80 90 100 TMRM (nm) pro-b large pre-b small pre-b 0 1 2 4 8 16 32 64 128 256 CCCP (mm)
Supplemental Table I.
Supplemental Table I Male / Mean ± SEM n Mean ± SEM n Body weight, g 29.2±0.4 17 29.7±0.5 17 Total cholesterol, mg/dl 534.0±30.8 17 561.6±26.1 17 HDL-cholesterol, mg/dl 9.6±0.8 17 10.1±0.7 17 Triglycerides,
SUPPLEMENTARY METHODS
SUPPLEMENTARY METHODS Histological analysis. Colonic tissues were collected from 5 parts of the middle colon on day 7 after the start of DSS treatment, and then were cut into segments, fixed with 4% paraformaldehyde,
Nature Immunology doi: /ni.3268
Supplementary Figure 1 Loss of Mst1 and Mst2 increases susceptibility to bacterial sepsis. (a) H&E staining of colon and kidney sections from wild type and Mst1 -/- Mst2 fl/fl Vav-Cre mice. Scale bar,
Supplementary Figure 1. IL-12 serum levels and frequency of subsets in FL patients. (A) IL-12
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Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained
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Supplemental Figure 1
Supplemental Figure 1 1a 1c PD-1 MFI fold change 6 5 4 3 2 1 IL-1α IL-2 IL-4 IL-6 IL-1 IL-12 IL-13 IL-15 IL-17 IL-18 IL-21 IL-23 IFN-α Mut Human PD-1 promoter SBE-D 5 -GTCTG- -1.2kb SBE-P -CAGAC- -1.kb
SUPPLEMENT Supplementary Figure 1: (A) (B)
SUPPLEMENT Supplementary Figure 1: CD4 + naïve effector T cells (CD4 effector) were labeled with CFSE, stimulated with α-cd2/cd3/cd28 coated beads (at 2 beads/cell) and cultured alone or cocultured with
Supplemental Information. CD4 + CD25 + Foxp3 + Regulatory T Cells Promote. Th17 Cells In Vitro and Enhance Host Resistance
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a b c Esophageal eosinophilia
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Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice
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Type of file: PDF Title of file for HTML: Supplementary Information Description: Supplementary Figures
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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
Supplementary Figures
Supplementary Figures Supplementary Fig. 1. Surface thiol groups and reduction of activated T cells. (a) Activated CD8 + T-cells have high expression levels of free thiol groups on cell surface proteins.
Supplementary information to: Mechanism of lipopolysaccharide-induced skin edema formation in the mouse
Supplementary information to: Mechanism of lipopolysaccharide-induced skin edema formation in the mouse Ivan Zanoni*, Renato Ostuni*, Simona Barresi, Marco Di Gioia, Achille Broggi, Barbara Costa, Roberta
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Supplementary Figure 1. Deletion of Smad3 prevents B16F10 melanoma invasion and metastasis in a mouse s.c. tumor model.
A B16F1 s.c. Lung LN Distant lymph nodes Colon B B16F1 s.c. Supplementary Figure 1. Deletion of Smad3 prevents B16F1 melanoma invasion and metastasis in a mouse s.c. tumor model. Highly invasive growth
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Pearson r = P (one-tailed) = n = 9
8F4-Specific Lysis, % 1 UPN1 UPN3 8 UPN7 6 Pearson r =.69 UPN2 UPN5 P (one-tailed) =.192 4 UPN8 n = 9 2 UPN9 UPN4 UPN6 5 1 15 2 25 8 8F4, % Max MFI Supplementary Figure S1. AML samples UPN1-UPN9 show variable
<10. IL-1β IL-6 TNF + _ TGF-β + IL-23
3 ns 25 ns 2 IL-17 (pg/ml) 15 1 ns ns 5 IL-1β IL-6 TNF