Journal:

Article Title: Retrovirus Vectors Bearing Jaagsiekte Sheep Retrovirus Env Transduce Human Cells by Using a New Receptor Localized to Chromosome 3p21.3

doi:

Figure Lengend Snippet: Host range of LAPSN vector produced by JSRV-pseudotype packaging cells

Article Snippet: Mammalian cells, including SSF-123 primary sheep skin fibroblasts (gift from William Osborne, University of Washington, Seattle), HT-1080 human fibrosarcoma cells (American Type Culture Collection [ATCC] cell line CCL-121), 293 human kidney epithelial cells (ATCC CRL 1573), IB3 immortalized human bronchial epithelial cells ( 62 ), HeLa cervical carcinoma cells (ATCC CCL-2), NIH 3T3 thymidine kinase-deficient mouse embryo fibroblasts ( 60 ), Mus dunni tail fibroblasts ( 11 ), D17 canine osteosarcoma cells (ATCC CRL-6248), 208F rat embryo fibroblasts ( 51 ), MDBK bovine kidney epithelial cells (ATCC CCL-22), Vero African green monkey kidney epithelial cells (ATCC CCL-81), MF-NAN primary mouse (BALB/c) fibroblasts, MF-H1 primary mouse (C57BL/6) fibroblasts, and RbTE rabbit tracheal epithelial cells (gifts from Christine Halbert, Fred Hutchinson Cancer Research Center), were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (Hyclone).

Techniques: Plasmid Preparation, Produced

Journal: Biomolecules

Article Title: Endothelial-Cell-Derived Human Secretory Leukocyte Protease Inhibitor (SLPI) Protects Cardiomyocytes against Ischemia/Reperfusion Injury

doi: 10.3390/biom9110678

Figure Lengend Snippet: Schematic diagram of the experimental design. Vascular endothelial cells (EC) (EA.hy926) were stably transfected by a eukaryotic expression vector containing human secretory leukocyte protease inhibitor (SLPI) mRNA ( hspli ) ( A ). The stable cells overexpressing recombinant human SLPI were used for further experiments. Determination of the paracrine effect of endothelial-derived SLPI on cardiomyocyte (CM) (H9c2) subjected to hypoxia/reoxygenation (H/R) injury was performed via techniques following two different principles: co-culture between SLPI-overexpressing EA.hy926 cells and H9c2 cells by the Transwell culture system ( B ), and conditioned medium from SLPI-overexpressing EA.hy926 cells ( C ).

Article Snippet: The human umbilical vein cell line (EA.hy926) and rat embryonic cardiomyocyte cell line (H9c2) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) as ATCC-CRL2992 and ATCC-CRL1446, respectively.

Techniques: Stable Transfection, Transfection, Expressing, Plasmid Preparation, Protease Inhibitor, Recombinant, Derivative Assay, Co-Culture Assay

Journal: Biomolecules

Article Title: Endothelial-Cell-Derived Human Secretory Leukocyte Protease Inhibitor (SLPI) Protects Cardiomyocytes against Ischemia/Reperfusion Injury

doi: 10.3390/biom9110678

Figure Lengend Snippet: The effect of rhSLPI secreted from the endothelial cell line protected the cardiomyocyte cell line from hypoxia/reoxygenation. ( A ) Wild-type (EA-WT) and SLPI-overexpressing (EA-SLPI) EA.hy926 cells were co-cultured with the H9c2 cell line at ratios 1:1 and 1:3, then subjected to H/R, and the cell viability was determined by MTT; ( B ) The culture media from wild-type (EA-WT) and SLPI-overexpressing (EA-SLPI) EA.hy926 cells were transferred to culture H9c2 cells at ratios of 1:1 and 1:3; then, the cells were exposed to H/R and the cell viability was determined by MTT. After transferring the media from wild-type (EA-WT) and SLPI-overexpressing (EA-SLPI) EA.hy926 cells at ratios of 1:1 and 1:3 to the H9c2 cells, ( C ) the cells were exposed to H2DCFDA followed by H/R, and the intracellular ROS levels were determined by spectrophotometry. Each bar represents the mean ± SEM. * p < 0.05 vs. the H9c2 not subjected to H/R (ANOVA), # p < 0.05 vs. the H9c2 subjected to H/R (ANOVA), † p < 0.05 vs. the 1:1 or 1:3 culture ratio group (ANOVA, n =3).

Article Snippet: The human umbilical vein cell line (EA.hy926) and rat embryonic cardiomyocyte cell line (H9c2) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) as ATCC-CRL2992 and ATCC-CRL1446, respectively.

Techniques: Cell Culture, Transferring, Spectrophotometry

Journal: Biomolecules

Article Title: Endothelial-Cell-Derived Human Secretory Leukocyte Protease Inhibitor (SLPI) Protects Cardiomyocytes against Ischemia/Reperfusion Injury

doi: 10.3390/biom9110678

Figure Lengend Snippet: Determination of the effect of rhSLPI secreted from endothelial cells on the cardiomyocyte cell line subjected to H/R in terms of the cellular signaling response. The media from cultured wild-type (EA-WT) and SLPI-overexpressing (EA-SLPI) EA.hy926 cells were transferred to culture H9c2 cells; then, the cells were exposed to H/R, and the activation of Akt ( A ) and p-38 MAPK ( B ) was determined by Western blot analysis. Each bar represents the phosphorylation ratio (phospho/total) of Akt and p38 MAPK. * p < 0.05 vs. H9c2 not subjected to H/R (ANOVA), # p < 0.05 vs. H9c2 subjected to H/R (ANOVA), † p < 0.05 vs. the H9c2/EA-WT medium group (ANOVA, n = 3).

Article Snippet: The human umbilical vein cell line (EA.hy926) and rat embryonic cardiomyocyte cell line (H9c2) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) as ATCC-CRL2992 and ATCC-CRL1446, respectively.

Techniques: Cell Culture, Activation Assay, Western Blot, Phospho-proteomics

Journal: Biomolecules

Article Title: Endothelial-Cell-Derived Human Secretory Leukocyte Protease Inhibitor (SLPI) Protects Cardiomyocytes against Ischemia/Reperfusion Injury

doi: 10.3390/biom9110678

Figure Lengend Snippet: Determination of the effect of rhSLPI secreted from endothelial cells on the cardiomyocyte cell line subjected to H/R in terms of the apoptosis pathway. The media from cultured wild-type (EA-WT) and SLPI-overexpressing (EA-SLPI) EA.hy926 cells were transferred to culture H9c2 cells; then, the cells were exposed to H/R, and the apoptotic proteins, including Bax/Bcl-2 ( A ), caspase-3 ( B ), and caspase-8 ( C ), were determined by Western blot analysis. Each bar represents the ratio relative to the reference protein (GAPDH). * p < 0.05 vs. H9c2 not subjected to H/R (ANOVA), # p < 0.05 vs. H9c2 subjected to H/R (ANOVA, n = 3).

Article Snippet: The human umbilical vein cell line (EA.hy926) and rat embryonic cardiomyocyte cell line (H9c2) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) as ATCC-CRL2992 and ATCC-CRL1446, respectively.

Techniques: Cell Culture, Western Blot

Journal: Biomolecules

Article Title: Endothelial-Cell-Derived Human Secretory Leukocyte Protease Inhibitor (SLPI) Protects Cardiomyocytes against Ischemia/Reperfusion Injury

doi: 10.3390/biom9110678

Figure Lengend Snippet: A schematic diagram of the major findings in this study. The rhSLPI that was overexpressed and secreted from vascular endothelial cells could protect not only endothelial cells but also cardiomyocytes from I/R injury. The mechanisms for this could be due to the attenuation of intracellular ROS production, activation of Akt and p38 MAPK phosphorylation, and reduction of apoptotic regulatory proteins (Bcl-2, caspase-3, caspase-8).

Article Snippet: The human umbilical vein cell line (EA.hy926) and rat embryonic cardiomyocyte cell line (H9c2) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) as ATCC-CRL2992 and ATCC-CRL1446, respectively.

Techniques: Activation Assay, Phospho-proteomics