Journal: Antioxidants & Redox Signaling

Article Title: NADPH Oxidase 4 Regulates Inflammation in Ischemic Heart Failure: Role of Soluble Epoxide Hydrolase

doi: 10.1089/ars.2018.7548

Figure Lengend Snippet: Cardiac remodeling and macrophage infiltration is reduced in Nox4−/− mice after LAD ligation. WT and Nox4−/− mice hearts were analyzed by immunofluorescence microscopy 2 weeks after sham or from the peri-infarct region after LAD ligation (A) Cardiomyocyte CSA was measured by WGA staining. (B) Capillary density was measured by vWF staining. (C) Cell death was measured in cardiomyocytes by co-staining with TUNEL and MYH7, a cardiomyocyte cell marker. (arrowheads indicate TUNEL+ nuclei). (D) Macrophages were identified by CD68 staining in sham-operated mice and in the border region of the infarct zone of LAD-ligated mice. MYH7 was used to identify the border region. (E) Quantification of cardiomyocyte CSA, (F) quantification of capillary density, (G) quantification of TUNEL+ cells, and (H) quantification of CD68+ cells. Data are the mean ± SEM, n = 9. Scale is 100 μm. *p < 0.05, **p < 0.01, and ****p < 0.0001. CSA, cross-sectional area; MYH7, myosin heavy chain beta; TUNEL, terminal deoxynucleotidyl transferase; vWF, von Willebrand Factor; WGA, wheat germ agglutinin.

Article Snippet: Cardiomyocyte CSA was measured by staining heart sections with WGA (Cat No. RL-1022; Vector Labs, Burlingame, CA), which stains cell membranes.

Techniques: Ligation, Immunofluorescence, Microscopy, Staining, TUNEL Assay, Marker

Journal: Antioxidants & Redox Signaling

Article Title: NADPH Oxidase 4 Regulates Inflammation in Ischemic Heart Failure: Role of Soluble Epoxide Hydrolase

doi: 10.1089/ars.2018.7548

Figure Lengend Snippet: Adverse left ventricular remodeling in human hearts with ICM. ICM hearts (n = 19) were compared with NF hearts (n = 24). (A) Representative images of left ventricular cross-sections were stained with WGA (top panels). Cardiomyocyte cross-sectional area was measured by using Image J (n ∼ 500–600 cells; middle panel). (B) Representative images of left ventricular cross-sections stained with von Willebrand factor to determine capillary density (top panel). Quantification of capillary density (n ∼ 100–125 microscopic fields; middle panel) (C) Representative fluorescent microscopy images of TUNEL-stained left ventricular sections (arrowheads indicate TUNEL+ nuclei; upper panel). Quantification of TUNEL+ nuclei (n ∼ 100–125 cross-sectional areas, middle panel) (D) Representative images of left ventricular cross-sections stained with Verhoeff sirius red (upper panel), and collagen was quantified by using Image J (middle panel). Data are the mean ± SEM. Scale bar is 100 μm. Scatter plots showing correlation between NOX4 protein levels and specific left ventricular remodeling markers in all human heart samples (bottom panels). The regression lines in each graph show the 95% confidence intervals (dotted lines), and R2 values are shown for each plot. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Cardiomyocyte CSA was measured by staining heart sections with WGA (Cat No. RL-1022; Vector Labs, Burlingame, CA), which stains cell membranes.

Techniques: Staining, Microscopy, TUNEL Assay

Journal: BMC Pharmacology & Toxicology

Article Title: Interaction between AhR and HIF-1 signaling pathways mediated by ARNT/HIF-1β

doi: 10.1186/s40360-022-00564-8

Figure Lengend Snippet: BaP in A549 cells enhanced the FRET fluorescence signal of pAcGFP1-N1-AhR and pDsRed-Monomer-ARNT. A pAcGFP1-N1-AhR and pDsRed-Monomer-ARNT recombinant plasmids were transfected into A5F49 cells. After 24 h, the cells were treated with 0, 2, 4, and 8 μM BaP, and then the following channels were used for high content detection: nucleus, donor: AhR, receptor: ARNT and FRET. B Bar chart showing changes in the ratios of FRET signal strength,* P < 0.05, ** P < 0.01, n = 3

Article Snippet: The chemicals used were purchased from the following companies: RPMI-1640 medium (Hyclone, Logan, UT, USA), fetal bovine serum (Gibco, Grand Island, NY, USA), HIF-1α stabilizing regent CoCl 2 and AhR ligand BaP (Sigma-Aldrich, St. Louis, MO, USA), pDsRed-Monomer-N1 expression vector and pAcGFP1-N1 expression vector (Clontech, Mountain View, CA, USA), Revert Aid First Strand cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA, Cat. K1621), BglII, KpnI enzyme (Takara, Shiga, Japan), Hoechst 33342(Thermo Fisher Scientific, Waltham, MA, USA, Cat. 1022) CAIX antibody (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-365,900), CYP1B1 antibody (Abcam, Cambridge, UK, Cat. ab33586), CYP1A1 antibody (Abcam, Cambridge, UK, Cat. ab3568), VEGF antibody (Abcam, Cambridge, UK, Cat. ab32152), HIF-1α antibody (Abcam, Cambridge, UK, Cat. ab1), and AhR antibody (Abcam, Cambridge, UK, Cat. ab84833), HIF-1β/ARNT antibody (Cell Signaling Technology, Danvers, MA, USA, Cat. 3781S).

Techniques: Fluorescence, Recombinant, Transfection

Journal: BMC Pharmacology & Toxicology

Article Title: Interaction between AhR and HIF-1 signaling pathways mediated by ARNT/HIF-1β

doi: 10.1186/s40360-022-00564-8

Figure Lengend Snippet: CoCl 2 in A549 cells reduced the FRET fluorescence signal of pAcGFP1-AhR and pDsRed-Monomer-ARNT. A pAcGFP1-N1-AhR and pDsRed-Monomer-ARNT recombinant plasmids were transfected into A549 cells; 24 h later, the cells were treated with 8 μM BaP for 24 h. B pAcGFP1-AhR and pDsRed-Monomer-ARNT recombinant plasmids were transfected into A549 cells; 24 h later, the cells were treated for 24 h with 8 μM BaP under hypoxic conditions. In A549 cells, BaP enhanced the pAcGFP1-AhR and pDsRed-Monomer-ARNT FRET fluorescent signal in dose-dependent manner,* P < 0.05, ** P < 0.01, n = 3

Article Snippet: The chemicals used were purchased from the following companies: RPMI-1640 medium (Hyclone, Logan, UT, USA), fetal bovine serum (Gibco, Grand Island, NY, USA), HIF-1α stabilizing regent CoCl 2 and AhR ligand BaP (Sigma-Aldrich, St. Louis, MO, USA), pDsRed-Monomer-N1 expression vector and pAcGFP1-N1 expression vector (Clontech, Mountain View, CA, USA), Revert Aid First Strand cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA, Cat. K1621), BglII, KpnI enzyme (Takara, Shiga, Japan), Hoechst 33342(Thermo Fisher Scientific, Waltham, MA, USA, Cat. 1022) CAIX antibody (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-365,900), CYP1B1 antibody (Abcam, Cambridge, UK, Cat. ab33586), CYP1A1 antibody (Abcam, Cambridge, UK, Cat. ab3568), VEGF antibody (Abcam, Cambridge, UK, Cat. ab32152), HIF-1α antibody (Abcam, Cambridge, UK, Cat. ab1), and AhR antibody (Abcam, Cambridge, UK, Cat. ab84833), HIF-1β/ARNT antibody (Cell Signaling Technology, Danvers, MA, USA, Cat. 3781S).

Techniques: Fluorescence, Recombinant, Transfection

Journal: Developmental cell

Article Title: YAP Partially Reprograms Chromatin Accessibility to Directly Induce Adult Cardiogenesis in Vivo

doi: 10.1016/j.devcel.2019.01.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Phospho-YAP(Ser127) antibody Cell signaling technology Cat#4911 Rabbit YAP1 antibody Novus biologicals Cat#NB110-583538 Donkey anti-Rabbit, Alexa 488 Thermo Fisher Scientific Cat#A-21206 Donkey anti-Rabbit, Alexa 647 Thermo Fisher Scientific Cat#A-31573 Donkey anti-Rabbit, Alexa 546 Thermo Fisher Scientific Cat#A-10040 Donkey anti-Mouse, Alexa 488 Thermo Fisher Scientific Cat#A-21202 Donkey anti-Mouse, Alexa 546 Thermo Fisher Scientific Cat#A-10036 Donkey anti-chicken, Alexa 488 Thermo Fisher Scientific Cat#A-11039 Donkey anti-rat, Alexa 488 Thermo Fisher Scientific Cat#A-11006 IRDye ® 800CW Goat anti-Mouse IgG LI-COR Cat#925-32210 IRDye ® 680RD Goat anti-Rabbit IgG LI-COR Cat#925-68071 Mouse anti-cTnT Thermo Fisher Scientific Cat#MA5-12960 Mouse anti-cTnT-Alexa 647 conjugate BD Pharmingen cat#565744 Mouse anti-AuroraKB Novus Cat#NBP2-50039 Rat anti-PHH3 Abcam Cat#ab10543 Mouse anti-M2Flag Sigma-Aldrich Cat#F1804 Mouse anti-GAPDH Millipore Cat#CB1001 Rabbit Anti-PCM-1 Sigma-Aldrich Cat#HPA023370 Chicken anti-β-gal Abcam Cat#ab9361 Rabbit anti-N-cadherin Abcam Cat#ab76057 Rat anti-BrdU Accurate Chemical & Scientific corp Cat#OBT0030 Rabbit DYKDDDDK Tag Antibody Cell signaling technology Cat#2368 Bacterial and Virus Strains pCMV-flag YAP25SA Addgene, Kun-Liang Guan plasmid # 27371 Chemicals, Peptides, and Recombinant Proteins Tamoxifen Sigma-Aldrich Cat#T5648 Collagenase A Sigma-Aldrich Cat#10103586001 Blebbistatin Sigma-Aldrich Cat#B0560 di-4-ANEPS Invitrogen Cat#D-1199 BglII NEB Cat#R0143S EcoRI-HF® NEB Cat#R3101 BsrGI-HF® NEB Cat#R3575L T4 DNA ligase (3C) NEB Cat#R3575L Proteinase K Sigma-Aldrich Cat#P2308-25MG RNAse A Sigma-Aldrich Cat#10109142001 Protein-G Dynabeads ThermoFisher Scientific Cat#10004D Rhodamine conjugated WGA Vector Labs Cat#RL-1022 DAPI Thermo Fisher Scientific Cat#62248 OptiPrem Density Gradient Medium Sigma-Aldrich Cat#D1556 Critical Commercial Assays Click-iT EdU Alexa Fluor 647 Imaging Kit ThermoFisher Scientific Cat# {"type":"entrez-nucleotide","attrs":{"text":"C10340","term_id":"1535411","term_text":"C10340"}} C10340 iTaq ™ Universal SYBR ® Green Supermix Bio-Rad Cat#1725120 DpnII NEB Cat#R0543M T4 DNA ligase (4C) Roche Cat#10799009001 Proteinase K (4C) Thermo Fisher Cat#EO0491 NucleoMag 96 PCR kit Macherey-Nagel Cat#744100.1 Csp6I Thermo Fisher Cat#ER0211 Expand Long template polymerase Roche Cat#11681842001 Agencourt AMPure XP beads Beckman Coulter Cat#A63881 Tissue-Tek ® O.C.T.

Techniques: Plasmid Preparation, Recombinant, Imaging, SYBR Green Assay, Expressing, Software, Next-Generation Sequencing

Journal: Developmental cell

Article Title: YAP Partially Reprograms Chromatin Accessibility to Directly Induce Adult Cardiogenesis in Vivo

doi: 10.1016/j.devcel.2019.01.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rhodamine-conjugated WGA used was from Vector labs Cat#RL-1022.

Techniques: Plasmid Preparation, Recombinant, Imaging, SYBR Green Assay, Expressing, Software, Next-Generation Sequencing

Journal: BMC Pharmacology & Toxicology

Article Title: Interaction between AhR and HIF-1 signaling pathways mediated by ARNT/HIF-1β

doi: 10.1186/s40360-022-00564-8

Figure Lengend Snippet: Schematic diagram of eukaryotic expression vector. A Bioassay based on FRET. B pAcGFP1-AhR and pDsRed-Monomer-ARNT enzyme-digested results of agarose gel electrophoresis. C Recombinant plasmid transfection

Article Snippet: The chemicals used were purchased from the following companies: RPMI-1640 medium (Hyclone, Logan, UT, USA), fetal bovine serum (Gibco, Grand Island, NY, USA), HIF-1α stabilizing regent CoCl 2 and AhR ligand BaP (Sigma-Aldrich, St. Louis, MO, USA), pDsRed-Monomer-N1 expression vector and pAcGFP1-N1 expression vector (Clontech, Mountain View, CA, USA), Revert Aid First Strand cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA, Cat. K1621), BglII, KpnI enzyme (Takara, Shiga, Japan), Hoechst 33342(Thermo Fisher Scientific, Waltham, MA, USA, Cat. 1022) CAIX antibody (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-365,900), CYP1B1 antibody (Abcam, Cambridge, UK, Cat. ab33586), CYP1A1 antibody (Abcam, Cambridge, UK, Cat. ab3568), VEGF antibody (Abcam, Cambridge, UK, Cat. ab32152), HIF-1α antibody (Abcam, Cambridge, UK, Cat. ab1), and AhR antibody (Abcam, Cambridge, UK, Cat. ab84833), HIF-1β/ARNT antibody (Cell Signaling Technology, Danvers, MA, USA, Cat. 3781S).

Techniques: Expressing, Plasmid Preparation, Agarose Gel Electrophoresis, Recombinant, Transfection

Journal: BMC Pharmacology & Toxicology

Article Title: Interaction between AhR and HIF-1 signaling pathways mediated by ARNT/HIF-1β

doi: 10.1186/s40360-022-00564-8

Figure Lengend Snippet: BaP in A549 cells enhanced the FRET fluorescence signal of pAcGFP1-N1-AhR and pDsRed-Monomer-ARNT. A pAcGFP1-N1-AhR and pDsRed-Monomer-ARNT recombinant plasmids were transfected into A5F49 cells. After 24 h, the cells were treated with 0, 2, 4, and 8 μM BaP, and then the following channels were used for high content detection: nucleus, donor: AhR, receptor: ARNT and FRET. B Bar chart showing changes in the ratios of FRET signal strength,* P < 0.05, ** P < 0.01, n = 3

Article Snippet: The chemicals used were purchased from the following companies: RPMI-1640 medium (Hyclone, Logan, UT, USA), fetal bovine serum (Gibco, Grand Island, NY, USA), HIF-1α stabilizing regent CoCl 2 and AhR ligand BaP (Sigma-Aldrich, St. Louis, MO, USA), pDsRed-Monomer-N1 expression vector and pAcGFP1-N1 expression vector (Clontech, Mountain View, CA, USA), Revert Aid First Strand cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA, Cat. K1621), BglII, KpnI enzyme (Takara, Shiga, Japan), Hoechst 33342(Thermo Fisher Scientific, Waltham, MA, USA, Cat. 1022) CAIX antibody (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-365,900), CYP1B1 antibody (Abcam, Cambridge, UK, Cat. ab33586), CYP1A1 antibody (Abcam, Cambridge, UK, Cat. ab3568), VEGF antibody (Abcam, Cambridge, UK, Cat. ab32152), HIF-1α antibody (Abcam, Cambridge, UK, Cat. ab1), and AhR antibody (Abcam, Cambridge, UK, Cat. ab84833), HIF-1β/ARNT antibody (Cell Signaling Technology, Danvers, MA, USA, Cat. 3781S).

Techniques: Fluorescence, Recombinant, Transfection

Journal: BMC Pharmacology & Toxicology

Article Title: Interaction between AhR and HIF-1 signaling pathways mediated by ARNT/HIF-1β

doi: 10.1186/s40360-022-00564-8

Figure Lengend Snippet: CoCl 2 in A549 cells reduced the FRET fluorescence signal of pAcGFP1-AhR and pDsRed-Monomer-ARNT. A pAcGFP1-N1-AhR and pDsRed-Monomer-ARNT recombinant plasmids were transfected into A549 cells; 24 h later, the cells were treated with 8 μM BaP for 24 h. B pAcGFP1-AhR and pDsRed-Monomer-ARNT recombinant plasmids were transfected into A549 cells; 24 h later, the cells were treated for 24 h with 8 μM BaP under hypoxic conditions. In A549 cells, BaP enhanced the pAcGFP1-AhR and pDsRed-Monomer-ARNT FRET fluorescent signal in dose-dependent manner,* P < 0.05, ** P < 0.01, n = 3

Article Snippet: The chemicals used were purchased from the following companies: RPMI-1640 medium (Hyclone, Logan, UT, USA), fetal bovine serum (Gibco, Grand Island, NY, USA), HIF-1α stabilizing regent CoCl 2 and AhR ligand BaP (Sigma-Aldrich, St. Louis, MO, USA), pDsRed-Monomer-N1 expression vector and pAcGFP1-N1 expression vector (Clontech, Mountain View, CA, USA), Revert Aid First Strand cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA, Cat. K1621), BglII, KpnI enzyme (Takara, Shiga, Japan), Hoechst 33342(Thermo Fisher Scientific, Waltham, MA, USA, Cat. 1022) CAIX antibody (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-365,900), CYP1B1 antibody (Abcam, Cambridge, UK, Cat. ab33586), CYP1A1 antibody (Abcam, Cambridge, UK, Cat. ab3568), VEGF antibody (Abcam, Cambridge, UK, Cat. ab32152), HIF-1α antibody (Abcam, Cambridge, UK, Cat. ab1), and AhR antibody (Abcam, Cambridge, UK, Cat. ab84833), HIF-1β/ARNT antibody (Cell Signaling Technology, Danvers, MA, USA, Cat. 3781S).

Techniques: Fluorescence, Recombinant, Transfection