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1 Supplementary legends Supplemental figure S1. Apelin-TAMRA is functional and induces apelin receptor internalization. HEK-293T cells transiently expressing YFP tagged APJ were incubated for 1 hour with: no drug (Unstimulated) or 100nM Apelin-TAMRA (Stimulated). Representative images of each condition showing native YFP fluorescence (left panels, green), TAMRA fluorescence (middle left panels, red), DAPI (middle right panels, blue) and Merge (right panels). Representative pictures of each condition are shown (n=4). Bars = 10 µm.
2 Supplemental figure S2. Apelin modulates duodenal contractions. (A) Representative recording of duodenal mechanical contractions in response to Krebs-Ringer (Control) or Apelin 100nM. (B) Ex vivo measurement of duodenal mechanical contractions amplitude in response to Krebs-Ringer (Control), L-NMMA or Iso (Isoproterenol). n=5-6 per group. **P<0.01, ***P<0,001. (C) Ex vivo measurement of duodenal mechanical contractions frequency before injection (Control Before) and in response to Krebs-Ringer (Control), increasing concentrations of Apelin (1pM-10µM), L-NMMA, Iso (Isoproterenol) and F13A alone or F13A plus Apelin (100nM or 1µM). n=4-6 per group. (D) Ex vivo measurement of jejunal (left panel) and ileal (right panel) mechanical contractions amplitude in response to Krebs-Ringer (Control) or increasing concentrations of Apelin (10pM-1µM). n=4-7 per group. (E) Ex vivo measurement of duodenal mechanical contractions amplitude in response to H20 (Control), Apelin 100nM (left panel) or 1µM (right panel), F13A or combination of Apelin plus F13A. n=5-6 per group. *P<0.05, **P<0.01.
3 Supplemental figure S3. Apelin acts on the ENS to modulate duodenal contractions. (A) Ex vivo measurement of duodenal acetylcholine release in basal condition and in response to Krebs-Ringer (Control) or increasing concentrations of Apelin (1pM-10µM). n = 5-13 per group. *P < 0.05, **P < 0.01, ***P < vs. Basal. (B) Duodenal NO release in response to Krebs-Ringer (Control), Apelin 100nM (left panel) or 1µM (right panel), F13A or combination of Apelin plus F13A. n=5-8 per group. *P<0.05, **P<0.01, ***P<0.001 vs. Control. #P<0.05, ##P<0.01, ###P<0.001 vs. F13A. P<0.05, P<0.001 vs. F13A + Apelin 1µM. (C) Representative recording of duodenal electrical activity in response to Krebs-Ringer (Control) or Apelin 100nM. (D) In vivo measurement of duodenal electrical activity in response to H20 (Control), Apelin 100nM (left panel) or 1µM (right panel), F13A or combination of Apelin plus F13A. n=4-7 per group. Left panel: *P<0,001 vs. Control, vs. Apelin 100nM + F13A and vs. F13A. #P<0.001 vs. Control, vs. Apelin 100nM and vs. F13A. $P<0.001 vs. Control, vs. Apelin 100nM and vs. Apelin 100nM + F13A. Right panel: *P<0,01 vs. Control, vs. Apelin 1µM + F13A and vs. F13A. #P<0.01 vs. Control, vs. Apelin 1µM and vs. F13A. $P<0.01 vs. Control, vs. Apelin 1µM and vs. Apelin 1µM + F13A.
4 Supplemental figure S4. Duodenal apelin modulates NO release in the hypothalamus. (A) In vivo effect of intragastric perfusion of H20 (Control) or Isoproterenol (Iso) on NO hypothalamic release frequency (left panel) and amplitude (right panel). n=5-7 per group. *P<0.05 vs. Control. (B) In vivo effect of intragastric perfusion of H20 (Control), Apelin 100nM (left panel) or 1µM (right panel), F13A or combination of Apelin plus F13A. n=4-7 per group. **P<0,01, ***P<0,001. (C) In vivo effect of intragastric perfusion of H20 (Control), Apelin 100nM alone or with F13A, Apelin 1µM alone or with F13A, or F13A alone, on hypothalamic NO release amplitude. n=4-7 per group. (D) In vivo effect of intragastric perfusion of H20 (Control), Apelin 100nM (left panel) or 1µM (right panel), Hexamethonium 500µM (Hx 500µM) or combination of Apelin plus Hexamethonium 500µM, on hypothalamic NO release amplitude. n=5-8 per group. (E) In vivo effect of intragastric perfusion of H20 (Control) and Apelin 100nM in mice pretreated or not by Capsaicin on hypothalamic NO release amplitude. n=4-7 per group.
5 Supplemental figure S5. Hypothalamic NO mediates the effects of intestinal Apelin in muscle glucose utilization. In vivo measurement of glucose entry in liver (A), muscle (B) and adipose tissue (C) in response to oral gavage of radioactive glucose in combination with H20 (Control) or Apelin 1µM after an icv injection of acsf (Control) or L-NMMA. n=8-12 per group. **P<0.01.
6 Supplemental figure S6. (A) Ex vivo measurement of duodenal Apelin of everted duodenal sacs incubated in Krebs-Ringer with 10g/L of glucose alone (Control), or with Apelin 1µM. n=5 per group. *P<0.05 vs. Control. (B) In vivo measurement of duodenal, jejunal and ileal Apelin in response to oral gavage of glucose with H2O (Control) or Apelin 1µM. n=4 per group. *P < In vivo measurement of glucose entry in liver (C) and adipose tissue (D) in response to oral gavage of radioactive glucose, in mice orally gavaged for one week with H20 (Control) or Apelin 1µM. n=7 per group. *P<0.05 vs. Control.
7 Supplemental figure S7. Relative expression of Apelin (A) and APJ (B) mrna in duodenum, jejunum and ileum of NC or HFD mice. n=5-6 per group. **P<0.01. (C) Ex vivo measurement of duodenal mechanical contraction frequency before injection (Control HFD Before) and in response to Krebs- Ringer (Control HFD) or Apelin 1µM in HFD mice. n=5-6 per group. (D) In vivo effect of intragastric perfusion of H20 (Control HFD) or Apelin 1µM, on NO hypothalamic release amplitude in HFD mice. n=4-6 per group.
Supplementary Figure 1) GABAergic enhancement by leptin hyperpolarizes POMC neurons A) Representative recording samples showing the membrane
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Supplementary Figure 1: Co-localization of reconstituted L-PTC and dendritic cells
a CD11c Na + K + ATPase Na + K + ATPase CD11c x-y CD11c Na + K + ATPase Na + K + ATPase CD11c x-z c b x-y view BoNT NAPs CD11c BoNT CD11c NAPs BoNT NAPs CD11c 90 x-z view Apical Basolateral Supplementary
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Supplementary Figure 1
Supplementary Figure 1 Localization of virus injections. (a) Schematic showing the approximate center of AAV-DIO-ChR2-YFP injection sites in the NAc of Dyn-cre mice (n=8 mice, 16 injections; caudate/putamen,
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Supplemental Material Supplemental Figure Legends Fig. S1 A. Generation and characterization of Ras-expressing autophagycompetent and -deficient cell lines. HA-tagged H-ras V12 was stably expressed in
c Ischemia (30 min) Reperfusion (8 w) Supplementary Figure bp 300 bp Ischemia (30 min) Reperfusion (4 h) Dox 20 mg/kg i.p.
a Marker Ripk3 +/ 5 bp 3 bp b Ischemia (3 min) Reperfusion (4 h) d 2 mg/kg i.p. 1 w 5 w Sacrifice for IF size A subset for echocardiography and morphological analysis c Ischemia (3 min) Reperfusion (8
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Supplementary Figures Supplementary Figure 1. Development of the camp biosensor targeted to the SERCA2a microdomain.
Supplementary Figures Supplementary Figure 1. Development of the camp biosensor targeted to the SERCA2a microdomain. A B C (A) Schematic representation of the new constructs designed for local camp imaging.
Expression constructs
Gene expressed in bebe3 ZmBEa Expression constructs 35S ZmBEa Pnos:Hygromycin r 35S Pnos:Hygromycin r 35S ctp YFP Pnos:Hygromycin r B -1 Chl YFP- Merge Supplemental Figure S1: Constructs Used for the Expression
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DOI:.38/ncb3399 a b c d FSP DAPI 5mm mm 5mm 5mm e Correspond to melanoma in-situ Figure a DCT FSP- f MITF mm mm MlanaA melanoma in-situ DCT 5mm FSP- mm mm mm mm mm g melanoma in-situ MITF MlanaA mm mm
Capsaicin (3.3 mmol/l)
A 18 16 14 1 8 6 4 Vehicle BCTC (.8 mmol/l) Sensory deafferentation Time (min) Capsaicin (3.3 mmol/l) N=3 4 * P
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Supplemental data Supplemental Figure Legends Supplemental Figure 1. Supplemental Figure 2.
Supplemental data Supplemental Figure Legends Supplemental Figure 1. Analysis of deletion of AMPK!2 in POMC and AgRP neurons in control and POMC!2KO and AgRP!2KO mice. (A) mmunofluorescence analysis for
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1.5 ASK1KO fed. fasted 16 hrs w/o water. Fed. 4th. 4th WT ASK1KO N=29, 11(WT), ,5(ASK1KO) ASK1KO ASK1KO **** Time [h]
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7: 13: 19: 1: 7: 151117 a 151117 4th 4th b c RQ.95 KO.9.85.8.75.7 light dark light dark.65 7: 19: 7: 19: 7: Means ± SEM, N=6 RQ 1..9.8.7.6.6 KO CL (-) CL (+) ibat weight ratio (/body weight) [%].5.4.3.2.1
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Supplementary Figure 1
Supplementary Figure 1 Supplementary Figure 1 Schematic depiction of the tandem Fc GDF15. Supplementary Figure 2 Supplementary Figure 2 Gfral mrna levels in the brains of both wild-type and knockout Gfral
Supplementary Figure 1
Combination index (CI) Supplementary Figure 1 2. 1.5 1. Ishikawa AN3CA Nou-1 Hec-18.5...2.4.6.8 1. Fraction affected (Fa) Supplementary Figure 1. The synergistic effect of PARP inhibitor and PI3K inhibitor
glucagon receptor AgRP merged color map I corr = 0.76±0.024 glucagon receptor DAPI merged
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Supplementary Data SUPPLEMENTARY FIG. S1. Representative counting fields used in quantification of the in vitro neural differentiation of pattern of anpcs. A panel of lineage-specific markers were used
Supplementary Figure 1
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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
mm Distance (mm)
b a Magnet Illumination Coverslips MPs Objective 2575 µm 1875 µm 1575 µm 1075 µm 875 µm 545 µm 20µm 2 3 0.5 0.3mm 1 1000 100 10 1 0.1 1000 100 10 1 0.1 Field Induction (Gauss) 1.5 0 5 10 15 20 Distance
Supplementary Figure 1.
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Supplemental Table S. Tumorigenicity and metastatic potential of 44SQ cell subpopulations a Tumorigenicity b Average tumor volume (mm ) c Lung metastasis d CD high /4 8. 8/ CD low /4 6./ a Mice were injected
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eee Supplementary Figure 1 Hyphae induce NET release, but yeast do not. (a) NET release by human peripheral neutrophils stimulated with a hgc1 yeast-locked C. albicans mutant (yeast) or pre-formed WT C.
Supplementary Information
Supplementary Information Supplementary Figure 1. EBV-gB 23-431 mainly exists as trimer in HEK 293FT cells. (a) Western blotting analysis for DSS crosslinked FLAG-gB 23-431. HEK 293FT cells transfected
c-abl PDGFRα PDGFRβ VEGFR-1 VEGFR-2 FLT-3 c-fms c-kit Ref. 2 (ND) 9.4 (ND) Supplementary Table 2. Chemical structure of the RTKIs used in this study.
SUPPLEMETARY DATA Supplementary Table 1. Upper and lower entries in each cell represent IC50 (nm) values determined by a biochemical kinase assay or cellular tyrosine kinase phosphorylation, respectively.
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DOI: 10.1038/ncb2211 a! mir-143! b! mir-103/107! let-7a! mir-144! mir-122a! mir-126-3p! mir-194! mir-27a! mir-30c! Figure S1 Northern blot analysis of mir-143 expression dependent on feeding conditions.
Postn MCM Smad2 fl/fl Postn MCM Smad3 fl/fl Postn MCM Smad2/3 fl/fl. Postn MCM. Tgfbr1/2 fl/fl TAC
A Smad2 fl/fl Smad3 fl/fl Smad2/3 fl/fl Tgfbr1/2 fl/fl 1. mm B Tcf21 MCM Tcf21 MCM Smad3 fl/fl Tcf21 MCM Smad2/3 fl/fl Tcf21 MCM Tgfbr1/2 fl/fl αmhc MCM C 1. mm 1. mm D Smad2 fl/fl Smad3 fl/fl Smad2/3
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