Lines. Levels of ErbB3 protein were quantified using western blot analysis (see Material and Methods) by densitometry. The graph represents the relative ErbB3 expression in elisidepsin-sensitive (IC50#1 mM) and -resistant (IC50.1 mM) cell lines. The Mann-Whitney test showed a statistically significant p value of 0.015. (TIF) Figure S3 Elisidepsin cell sensitivity is associated withFigure S4 Generation and characterization of elisidepsin-resistant cell lines from colon and lung. A) Cells were lysed, proteins were extracted and western blots performed with an equal amount of cell lysate (50 mg protein). Expression of epithelial (E-cadherin, b-catenin, c-catenin)- and mesenchymal (vimentin, Slug, Snail, Twist)-associated proteins differentiates between elisidepsin-sensitive and elisidepsin-resistant cell lines. b-actin was used as an internal control. These western blots were performed in triplicate. B) Expression levels HER1, HER2, HER3, HER4, pAkt, and pMAPK were analyzed by western blot using 50 mg 18325633 of protein cell lysate. The membranes were stripped and reprobed with anti-b-actin to Title Loaded From File verify equal protein loading. HCT 116 (C) and A549 (D) elisidepsin-sensitive cancer cell lines were rendered resistant by persistent exposure to increasing concentrations of elisidepsin. Cells were treated with elisidepsin at the indicated concentrations for 72 h and cell viability was measured using a crystal violet assay. Error bars show the SD of three replicate experiments. C, control; R, resistance. (TIF) Figure SChemical structure of 24272870 elisidepsin.(TIF)AcknowledgmentsWe would like to thank Dr. Atanasio Pandiella for providing the HER3 antibody and for helpful discussions during the preparation of the manuscript.HER3 expression levels. Levels of HER1, HER2, HER3 and HER4 protein were quantified with western blot analysis (Fig. 4) and subsequent densitometry. Cells that have an elisidepsin IC50 value of #1 mM were considered sensitive to the drug. The graph represents the HER family members expression relative to elisidepsin sensitivity. A statistically significance relationship between HER3 expression levels and elisidepsin sensitivity was found (Mann-Whitney test: p = 0.0091) but not with the other members. (TIF)Author ContributionsConceived and designed the experiments: CT SRC JHL. Performed the experiments: CT RM. Analyzed the data: CT JHL. Contributed reagents/ materials/analysis tools: CT RM MA SRC JHL. Wrote the paper: CT JHL.
Cardiac muscle cells (cardiomyocytes) are frequently thought to be the most abundant cell type in the adult heart. However, multiple studies have shown that cardiac chamber walls comprise high numbers of non-myocyte cells. These cells and their milieu (the extracellular space between cardiomyocyte fibers) constitute the cardiac interstitium [1?]. Due to the small relative size of cardiac interstitial cells (CICs) and the enormous contribution of cardiomyocytes to cardiac mass, the Sense 59TGTGGGAATCCGACGAATG-39 and antisense 59- GTCATATGGTGGAGCTGTGGG-39 for N-Cadherin; sense 59CGGGAATGCAGTTGAGGATC-39 and proportion of CICs versus cardiac muscle cells in the heart is frequently underestimated. In this regard, recent reports suggest that CICs could represent up to a 65 of non-cardiomyocyte cells in the organ [1?]. The biomedical importance of CICs is illustrated by their massive involvement in the remodeling of cardiac ventricular walls after myocardial infarction, a phenomenon that is characterized by a progressive fibrosis [4]. This ventricular remodeling involves the initiation of an inflammatory response and the mobilization of CICs. Both phenomen.Lines. Levels of ErbB3 protein were quantified using western blot analysis (see Material and Methods) by densitometry. The graph represents the relative ErbB3 expression in elisidepsin-sensitive (IC50#1 mM) and -resistant (IC50.1 mM) cell lines. The Mann-Whitney test showed a statistically significant p value of 0.015. (TIF) Figure S3 Elisidepsin cell sensitivity is associated withFigure S4 Generation and characterization of elisidepsin-resistant cell lines from colon and lung. A) Cells were lysed, proteins were extracted and western blots performed with an equal amount of cell lysate (50 mg protein). Expression of epithelial (E-cadherin, b-catenin, c-catenin)- and mesenchymal (vimentin, Slug, Snail, Twist)-associated proteins differentiates between elisidepsin-sensitive and elisidepsin-resistant cell lines. b-actin was used as an internal control. These western blots were performed in triplicate. B) Expression levels HER1, HER2, HER3, HER4, pAkt, and pMAPK were analyzed by western blot using 50 mg 18325633 of protein cell lysate. The membranes were stripped and reprobed with anti-b-actin to verify equal protein loading. HCT 116 (C) and A549 (D) elisidepsin-sensitive cancer cell lines were rendered resistant by persistent exposure to increasing concentrations of elisidepsin. Cells were treated with elisidepsin at the indicated concentrations for 72 h and cell viability was measured using a crystal violet assay. Error bars show the SD of three replicate experiments. C, control; R, resistance. (TIF) Figure SChemical structure of 24272870 elisidepsin.(TIF)AcknowledgmentsWe would like to thank Dr. Atanasio Pandiella for providing the HER3 antibody and for helpful discussions during the preparation of the manuscript.HER3 expression levels. Levels of HER1, HER2, HER3 and HER4 protein were quantified with western blot analysis (Fig. 4) and subsequent densitometry. Cells that have an elisidepsin IC50 value of #1 mM were considered sensitive to the drug. The graph represents the HER family members expression relative to elisidepsin sensitivity. A statistically significance relationship between HER3 expression levels and elisidepsin sensitivity was found (Mann-Whitney test: p = 0.0091) but not with the other members. (TIF)Author ContributionsConceived and designed the experiments: CT SRC JHL. Performed the experiments: CT RM. Analyzed the data: CT JHL. Contributed reagents/ materials/analysis tools: CT RM MA SRC JHL. Wrote the paper: CT JHL.
Cardiac muscle cells (cardiomyocytes) are frequently thought to be the most abundant cell type in the adult heart. However, multiple studies have shown that cardiac chamber walls comprise high numbers of non-myocyte cells. These cells and their milieu (the extracellular space between cardiomyocyte fibers) constitute the cardiac interstitium [1?]. Due to the small relative size of cardiac interstitial cells (CICs) and the enormous contribution of cardiomyocytes to cardiac mass, the proportion of CICs versus cardiac muscle cells in the heart is frequently underestimated. In this regard, recent reports suggest that CICs could represent up to a 65 of non-cardiomyocyte cells in the organ [1?]. The biomedical importance of CICs is illustrated by their massive involvement in the remodeling of cardiac ventricular walls after myocardial infarction, a phenomenon that is characterized by a progressive fibrosis [4]. This ventricular remodeling involves the initiation of an inflammatory response and the mobilization of CICs. Both phenomen.
