Gallic acid prevents isoproterenol-induced cardiac hypertrophy and fibrosis through regulation of JNK2 signaling and Smad3 binding activity

Similar documents
hemodynamic stress. A. Echocardiographic quantification of cardiac dimensions and function 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.

E10.5 E18.5 P2 10w 83w NF1 HF1. Sham ISO. Bmi1. H3K9me3. Lung weight (g)

Protection against doxorubicin-induced myocardial dysfunction in mice by cardiac-specific expression of carboxyl terminus of hsp70-interacting protein

SUPPLEMENTAL MATERIAL. Supplementary Methods

Plasma exposure levels from individual mice 4 hours post IP administration at the

Fetal gene upregulation by 1-wk TAC is significantly increased in mice lacking RGS2.

Supporting Information

Serum cytokine levels in control and tumor-bearing male and female mice at day 15.

Tcf21 MCM ; R26 mtmg Sham GFP Col 1/3 TAC 8W TAC 2W. Postn MCM ; R26 mtmg Sham GFP Col 1/3 TAC 8W TAC 2W

Figure S1. Sorting nexin 9 (SNX9) specifically binds psmad3 and not psmad 1/5/8. Lysates from AKR-2B cells untreated (-) or stimulated (+) for 45 min

Supplementary Figure 1. Confocal immunofluorescence showing mitochondrial translocation of Drp1. Cardiomyocytes treated with H 2 O 2 were prestained

SUPPLEMENTARY INFORMATION

Control. csarnt -/- Cre, f/f

CYLD Negatively Regulates Transforming Growth Factor-β Signaling via Deubiquitinating Akt

p = formed with HCI-001 p = Relative # of blood vessels that formed with HCI-002 Control Bevacizumab + 17AAG Bevacizumab 17AAG

hexahistidine tagged GRP78 devoid of the KDEL motif (GRP78-His) on SDS-PAGE. This

Supplementary Information File

Supplemental Information

SUPPLEMENTARY INFORMATION

Supplementary Figure 1. Spatial distribution of LRP5 and β-catenin in intact cardiomyocytes. (a) and (b) Immunofluorescence staining of endogenous

In vivo bromodeoxyuridine (BrdU) incorporation was performed to analyze cell

a. b. c. d. e. f. g. h. i. j. k. l. m. n. o. p.

Supplementary Fig. S1. Schematic diagram of minigenome segments.

Supplementary Materials and Methods

SUPPLEMENTARY INFORMATION

doi: /nature14508 Rappsilber et al.

Supplemental Figure 1

BNP mrna expression in DR and DS rat left ventricles (n = 5). (C) Plasma norepinephrine

Supplement Material. chain (MLC2v) promoter to generate cardiac-specific MKK4 knockout mice (referred to

Figures S1-S5, Figure Legends, Table S1 List of primers used in the study

Postn MCM Smad2 fl/fl Postn MCM Smad3 fl/fl Postn MCM Smad2/3 fl/fl. Postn MCM. Tgfbr1/2 fl/fl TAC

Amniotic fluid stem cells provide considerable advantages in epidermal. regeneration: B7H4 creates a moderate inflammation

Supplementary Figure 1:

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14-

Nature Medicine: doi: /nm.4324

Probe. Hind III Q,!&#12?R'!! /0!!!!D1"?R'! vector. Homologous recombination

Supplementary Figure 1. PAQR3 knockdown inhibits SREBP-2 processing in CHO-7 cells CHO-7 cells were transfected with control sirna or a sirna

Supplementary Figure 1. Baf60c and baf180 are induced during cardiac regeneration in zebrafish. RNA in situ hybridization was performed on paraffin

Intracellular MHC class II molecules promote TLR-triggered innate. immune responses by maintaining Btk activation

Supplementary Figure 1: Expression of NFAT proteins in Nfat2-deleted B cells (a+b) Protein expression of NFAT2 (a) and NFAT1 (b) in isolated splenic

Supplementary Figure 1. Expression of CUGBP1 in non-parenchymal liver cells treated with TGF-β

Research article. Department of Pharmacology and Toxicology, Medical College of Georgia, Georgia Health Sciences University, Augusta, Georgia, USA.

Supplementary Table 1. The primers used for quantitative RT-PCR. Gene name Forward (5 > 3 ) Reverse (5 > 3 )

Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured

Supplementary Figures Supplementary Figure 1. Development of the camp biosensor targeted to the SERCA2a microdomain.

Supplementary Materials for

condition. Left panel, the HCT-116 cells were lysed with RIPA buffer containing 0.1%

AP VP DLP H&E. p-akt DLP

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION

ANGPTL2 increases bone metastasis of breast cancer cells through. Tetsuro Masuda, Motoyoshi Endo, Yutaka Yamamoto, Haruki Odagiri, Tsuyoshi

[Billing Code: ] DEPARTMENT OF HEALTH AND HUMAN SERVICES. Office of the Secretary. Findings of Research Misconduct

Appendix Table of Contents. 1. Appendix Figure legends S1-S13 and Appendix Table S1 and S2. 2. Appendix Figures S1-S13

Gene expression was analyzed by real time PCR (SYBR GREEN PCR Master. Mix, Roche Applied Science) using specific oligonucleotides.

SUPPLEMENTARY INFORMATION

Supplementary Figure 1.TRIM33 binds β-catenin in the nucleus. a & b, Co-IP of endogenous TRIM33 with β-catenin in HT-29 cells (a) and HEK 293T cells

Supplementary Materials for

Nature Genetics: doi: /ng Supplementary Figure 1. Parameters and consequences of mononuclear cardiomyocyte frequency.

SUPPLEMENTARY INFORMATION

Supplementary Figure 1 ITGB1 and ITGA11 increase with evidence for heterodimers following HSC activation. (a) Time course of rat HSC activation

Title: Cytosolic DNA-mediated, STING-dependent pro-inflammatory gene. Fig. S1. STING ligands-mediated signaling response in MEFs. (A) Primary MEFs (1

Supplementary Figure 1: si-craf but not si-braf sensitizes tumor cells to radiation.

Supplementary Information

SUPPLEMENTARY RESULTS

Control GST GST-RAP. α2-mg. 170 kda. b-actin. 42 kda LRP-1

BMP6 treatment compensates for the molecular defect and ameliorates hemochromatosis in Hfe knockout mice

Supplementary Materials for

Supplementary Table 1. List of primers used in this study

Table S1. Primer sequences used for qrt-pcr. CACCATTGGCAATGAGCGGTTC AGGTCTTTGCGGATGTCCACGT ACTB AAGTCCATGTGCTGGCAGCACT ATCACCACTCCGAAGTCCGTCT LCOR

Supplementary information

SUPPLEMENTARY FIGURES AND TABLE

TRAF6 ubiquitinates TGFβ type I receptor to promote its cleavage and nuclear translocation in cancer

GPR120 *** * * Liver BAT iwat ewat mwat Ileum Colon. UCP1 mrna ***

Supplementary data Supplementary Figure 1 Supplementary Figure 2

Figure S1. Reduction in glomerular mir-146a levels correlate with progression to higher albuminuria in diabetic patients.

ZL ZDF ZDF + E2 *** Visceral (g) ZDF

SUPPLEMENTARY INFORMATION

Programmed necrosis, not apoptosis, is a key mediator of cell loss and DAMP-mediated inflammation in dsrna-induced retinal degeneration

Minute TM Plasma Membrane Protein Isolation and Cell Fractionation Kit User Manual (v5)

Supplementary Materials

Supplementary Figures

Supplementary Figure 1. Western blot of hippocampal lysates from WT and Adcy1 KO mice demonstrates the specificity of the ADCY1 antibody.

5 -GGTAAAGCAGGTCTAGGTGGCTGACAGTCT-3. Cre-transgene allele was detected. by PCR using the primers; 5 -ACATGTTCAGGGATCGCCAG-3 and

Supplementary Figure 1: Co-localization of reconstituted L-PTC and dendritic cells

Figure 1. Effects of FGF21 on adipose tissue. (A) Representative histological. findings of epididymal adipose tissue (B) mrna expression of

Supplementary Materials for

Supplementary Figure 1. Repression of hepcidin expression in the liver of mice treated with

Supplementary Fig. 1. GPRC5A post-transcriptionally down-regulates EGFR expression. (a) Plot of the changes in steady state mrna levels versus

General Laboratory methods Plasma analysis: Gene Expression Analysis: Immunoblot analysis: Immunohistochemistry:

Fig. S1. Dose-response effects of acute administration of the β3 adrenoceptor agonists CL316243, BRL37344, ICI215,001, ZD7114, ZD2079 and CGP12177 at

Supplemental Data. TGF-β-mediated mir-181a expression promotes breast cancer metastasis by targeting Bim.

Supplementary Figure 1 Expression of Crb3 in mouse sciatic nerve: biochemical analysis (a) Schematic of Crb3 isoforms, ERLI and CLPI, indicating the

Atrogin-1/muscle atrophy F-box inhibits calcineurin-dependent cardiac hypertrophy by participating in an SCF ubiquitin ligase complex

Supplementary Figure 1

Tumour growth environment modulates Chk1 signalling pathways and sensitivity to Chk1 inhibition

Dipeptidyl Peptidase-4 Inhibitor Increases Vascular Leakage in Retina through VE-cadherin. Phosphorylation

Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk

Supplementary Figure 1. EC-specific Deletion of Snail1 Does Not Affect EC Apoptosis. (a,b) Cryo-sections of WT (a) and Snail1 LOF (b) embryos at

Genetic ablation of Acp1 (Lmptp) in mice prevents heart failure

Transcription:

Gallic acid prevents isoproterenol-induced cardiac hypertrophy and fibrosis through regulation of JNK2 signaling and Smad3 binding activity Yuhee Ryu 1,+, Li Jin 1,2+, Hae Jin Kee 1,, Zhe Hao Piao 3, Jae Yeong Cho 1, Gwi Ran Kim 1, Sin Young Choi 1, Ming Quan Lin 1,4, and Myung Ho Jeong 1, 1 Heart Research Center of Chonnam National University Hospital, Gwangju 51-757, Republic of Korea. 2 Jilin Hospital Affiliated with Jilin University, Jilin, China 3 The Second Hospital of Jilin University, Changchun, China 4 Yanbian University Hospital, 1327 Juzi Road, Yanbian, Jilin 133, China Supplementary Data 1

Supplementary Table 1 Supplementary Table 1. Primers for reverse transcription polymerase chain reaction (RT-PCR) and chromatin immunoprecipitation (ChIP) Gene (mouse) ANP BNP Collagen I Fibronectin Alpha SMA GAPDH (mouse) Collagen type I (ChIP) F, forward; R, reverse. Primer sequence (5ʹ to 3ʹ) F: TGGAGGAGAAGATGCCGGTAGAAGAT R: AGCGAGCAGAGCCCTCAGTTTGCT F: CTGAAGGTGCTGTCCCAGAT R: GTTCTTTTGTGAGGCCTTGG F: GAGCGGAGAGTACTGGATCG R: GCTTCTTTTCCTTGGGGTTC F: GATGCACCGATTGTCAACAG R: TGATCAGCATGGACCACTTC F: ACTGGGACGACATGGAAAAG R: AGAGGCATAGAGGGACAGCA F: GCATGGCCTTCCGTGTTCCT R: CCCTGTTGCTGTAGCCGTATTCAT F: TGAGAAGTGGCAGAGGAGGT R: GACTGCCACATCAAGGGTCT 2

2. Supplementary Figures Legends Supplementary Figure 1. Effect of gallic acid on isoproterenol (ISP)-treated H9c2 cells. H9c2 cells were incubated with ISP I (1 µmol/l) in the presence or absence of gallic acid (1 µmol/l). (A) Cells were fixed with 4% paraformaldehyde and incubated with sarcomeric α-actinin (1:2). Nuclei were stained with DAPI. Merged images are shown. (B) Cell size was measured. (C) Protein lysates were resolved using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with the indicated antibodies. Representative western blots are shown. (D-F) Quantification of β-mhc, ANP, and BNP protein levels was performed using densitometry. Protein expression was normalized to GAPDH. Values are means ± SD of 3 independent experiments. P < 5 versus vehicle. # P < 5 and ## P < 1 versus ISP. Supplementary Figure 2. Echocardiographic parameters in ISP-infused mice. (A) Representative M-mode images of left ventricles in mice infused with ISP (1~3 day) as indicated. (B F) Left ventricular posterior wall thickness, interventricular septum thickness, left ventricular end-systolic dimension, left ventricular end-systolic dimension, and fractional shortening were measured after ISP infusion. P < 5, P < 1, and P < 1 versus control. (G) Heart weight to body weight ratio. Supplementary Figure 3. Expression of MAPK signaling in ISP-treated cardiac fibroblasts. Rat cardiac fibroblasts were serum-starved and incubated with ISP I (1 µmol/l) at indicated time. Proteins were analyzed by western blot with anti-p-erk1/2 (Thr 22/Thy24), ERK, p-jnk1/2 (S423/425), and JNK antibodies. β-actin was used as an internal control. Supplementary Figure 4. Expression of Smad3 in ISP-treated cardiac fibroblasts. Gallic acid administration was started 2 week before infusion of ISP (3 day) in mice; sham + vehicle, ISP + vehicle, and ISP + gallic acid group. Proteins were subjected to western blotting with anti-smad3 antibody. GAPDH was used as a loading control. Smad3 protein was quantified by densitometry. # P<5 versus ISP + vehicle. NS indicates not significant. Supplementary Figure 5. Expression of phosphorylated Smad3 in ISP-treated cardiac fibroblasts. Rat cardiac fibroblasts were serum-starved and incubated with ISP I (1 µmol/l) at indicated time. Proteins were analyzed by western blot with anti-psmad3 (S423/S425) and Smad3 antibodies. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. 3

Supplementary Figure 1 A C Control + vehicle ISP + vehicle ISP + Gallic acid ISP - - - + + + + + + + + + Gallic acid - - - - - - 5 5 5 1 1 1 β-mhc ANP BNP GAPDH E Relative ANP protein 2. ## F Relative BNP protein B Cell size (arbitrary unit) D 6 4 2 Relative MHC protein 2. # ### ISP - + + Gallic acid - - + ## ISP - + + + Gallic acid - - 5 1 ISP - + + + Gallic acid - - 5 1 ISP - + + + Gallic acid - - 5 1

Supplementary Figure 2 A ISP d ISP 1 d ISP 2 d ISP 3 d B left ventricular posterior wall thickness (mm) C Interventricular septum thickness (mm) D left ventricular end-systolic dimension (mm) 4 3 2 1 E left ventricular end-diastolic dimension (mm) 6 4 2 F Fractional shortening (%) 8 6 4 2 G Heart weight / body weight ratio 6 4 2 n=12 n=2 n=2 n=13

Supplementary Figure 3 ISP Time (h) - 1 3 6 9 12 p-erk ERK p-jnk JNK β-actin Rat cardiac fibroblast

Supplementary Figure 4 Relative Smad3/GAPDH protein NS n=12 n=15 n=15 ISP - + + Gallic acid - - + #

Supplementary Figure 5 ISP (h) 1 3 6 9 12 p-smad3 (S423/425) Smad3 GAPDH Cardiac fibroblast 54 kda 54 kda 38 kda

2024 © healthdocbox.com Privacy Policy | Terms of Service | Feedback