Journal: PLoS ONE

Article Title: Expression of Beta-Defensin 131 Promotes an Innate Immune Response in Human Prostate Epithelial Cells

doi: 10.1371/journal.pone.0144776

Figure Lengend Snippet: ( A ) DEFB131 mRNA upregulation. RWPE-1 cells were transfected with an empty vector or the DEFB131-DDK-Myc vector. Quantitative real-time PCR analysis was used to determine mRNA levels. Expression levels were calculated from C T values and normalized to ACTB. Asterisks represent statistical significance at P < 0.0001. ( B ) Upregulation of DEFB131 protein. DEFB131 protein levels were analyzed by western blot in cell lysates. ( C ) Overexpression of secreted DEFB131 protein by western blot in culture supernatants. ( D, E ) Gene expression profiles for cytokines and chemokines in DEFB131-activated RWPE-1 cells were determined using quantitative real-time PCR analysis. Asterisks (* and **) represent statistical significance at P < 0.01 and P < 0.05.

Article Snippet: Cytokines and chemokines were measured using the Human Inflammatory Cytokines Multi-Analyte ELISArray kit (Qiagen) and the Human Common Chemokines Multi-Analyte ELISArray Kit (Qiagen) according to the manufacturer's instructions.

Techniques: Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Over Expression, Gene Expression

Journal: PLoS ONE

Article Title: Expression of Beta-Defensin 131 Promotes an Innate Immune Response in Human Prostate Epithelial Cells

doi: 10.1371/journal.pone.0144776

Figure Lengend Snippet: Supernatants obtained from RWPE-1 cells transfected with an empty vector or DEFB131-DDK-Myc were collected after 24 h. The levels of cytokines and chemokines released into the medium were quantified by multi-analyte ELISA. Error bars represent the standard error of the mean (SEM). Asterisks (* and **) represent statistical significance at P < 0.001 and P < 0.01.

Article Snippet: Cytokines and chemokines were measured using the Human Inflammatory Cytokines Multi-Analyte ELISArray kit (Qiagen) and the Human Common Chemokines Multi-Analyte ELISArray Kit (Qiagen) according to the manufacturer's instructions.

Techniques: Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Expression of Beta-Defensin 131 Promotes an Innate Immune Response in Human Prostate Epithelial Cells

doi: 10.1371/journal.pone.0144776

Figure Lengend Snippet: LTA binds to the TLR2/TLR6 receptor complex and activates AP1 and NF-κB transcription factors through stimulation of the p38MAPK and NF-κB pathways in RWPE-1 cells. Upon LTA stimulation, the activated transcription of AP1 and NF-κB markedly induces DEFB131 expression and secretion, followed by increased production of cytokines and chemokines. Secreted DEFB131 exhibits antimicrobial activity. DEFB131 induces cytokine and chemokine expression and subsequent immune cell recruitment. DEFB131 may contribute to the activation of prostate epithelial cell innate and adaptive immune responses.

Article Snippet: Cytokines and chemokines were measured using the Human Inflammatory Cytokines Multi-Analyte ELISArray kit (Qiagen) and the Human Common Chemokines Multi-Analyte ELISArray Kit (Qiagen) according to the manufacturer's instructions.

Techniques: Expressing, Activity Assay, Activation Assay