Journal: Journal of Leukocyte Biology

Article Title: Directed migration of mouse macrophages in vitro involves myristoylated alanine-rich C-kinase substrate (MARCKS) protein

doi: 10.1189/jlb.1211604

Figure Lengend Snippet: The MANS, but not the RNS, peptide inhibits cell migration. Cells were untreated or treated with 50 μM peptide prior to adding MCP-1 in a Transwell migration assay. Cells that migrated through the pores to the lower side of the insert membrane, were fixed and counted at 40× magnification. That number is given on each panel. Contrast was enhanced for this figure after the panels were assembled to clearly define the migrated cells from the visible membrane pores.

Article Snippet: MANS and RNS peptides were synthesized by Genemed Synthesis (San Antonio, TX, USA).

Techniques: Migration, Transwell Migration Assay, Membrane

Journal: Journal of Leukocyte Biology

Article Title: Directed migration of mouse macrophages in vitro involves myristoylated alanine-rich C-kinase substrate (MARCKS) protein

doi: 10.1189/jlb.1211604

Figure Lengend Snippet: J774A.1 cells were pretreated with 50 μM MANS or RNS. Migration was measured in response to: (A) MCP-1 (50 ng/ml); (B) MCP-1 (100 ng/ml); or (C) C5a (10 ng/ml). Cells were exposed for 3 h, at which time, migrating cells were quantified. Data represent mean ± sem; n = 3; **significantly different from media control; †significantly different from stimulated/media control; P < 0.05.

Article Snippet: MANS and RNS peptides were synthesized by Genemed Synthesis (San Antonio, TX, USA).

Techniques: Migration, Control

Journal: Journal of Leukocyte Biology

Article Title: Directed migration of mouse macrophages in vitro involves myristoylated alanine-rich C-kinase substrate (MARCKS) protein

doi: 10.1189/jlb.1211604

Figure Lengend Snippet: Isolated cells were pretreated with: (A) 25 μM or (B) 50 μM MANS or RNS peptide for 15 min. Migration was measured in response to MCP-1 (100 ng/ml). Cells were exposed for 3 h, at which time, migrating cells were quantified. Data represent mean ± sem; n = 3; **significantly different from media control; †significantly different from MCP-1/media control; P < 0.05.

Article Snippet: MANS and RNS peptides were synthesized by Genemed Synthesis (San Antonio, TX, USA).

Techniques: Isolation, Migration, Control

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) The panel shows the different mixtures of DNA (100 ng) and MMGP1 (1- 0.036; 2- 0.072; 3- 0.144; 4- 0.288; 5-0.576 µM; C-without peptide) run on a 1% agarose gel. (b) Quantitative assessment of unbound plasmid DNA to the peptide by gel densitometry analysis.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Agarose Gel Electrophoresis, Plasmid Preparation

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Measurement of intrinsic tryptophan fluorescence of MMGP1 on addition of plasmid DNA (b) Quenching of SYTO9 fluorescence in the presence of varying concentrations of MMGP1.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Fluorescence, Plasmid Preparation

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Treatment of DNA–MMGP1 complexes with proteinase K resulted in detection of plasmid DNA band on the agarose gel. C-Control SK+ plasmid DNA; 1-DNA–MMGP1 complex formed at 0.576 µM concentration of peptide; 2-DNA–MMGP1 complex treated with proteinase K (b) DNase I protection assay. The plasmid DNA (100 ng) was preincubated with the varying concentrations peptides such as, C+-no peptide; 1-0.576 µM; 2-0.288 µM; 3- 0.144 μM; 4-0.072 µM; 5-0.036 µM, respectively followed by treatment with DNase I. The DNase treated plasmid DNA was used as negative control (C-).

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Plasmid Preparation, Agarose Gel Electrophoresis, Control, Concentration Assay, Negative Control

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Expression of mouse β-actin gene in the presence of different concentrations of MMGP1 C-no peptide, 1-0.576 µM, 2-0.288 µM, 3- 0.144 μM, 4- 0.072; 5- 0.036 at 37 ° C for 1 h under in vitro condition. The transcribed product of 304 bases were analysed on 5% denaturing polyacrylamide gel. (b) Quantitative measurement of mouse β-actin gene expression in the presence of varying concentrations of MMGP1 by gel densitometry analysis.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Expressing, In Vitro, Gene Expression

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Confocal micrographs showing inhibition of transcription in C. albicans by MMGP1. The images are overlay of TMR-florescent azide (red), Hoechst 33342 (blue) and bright field micrographs of C. albicans cells. Intense EU staining (red fluorescence) was observed in nucleus after 2 h of treatment with MMGP1 and prolonged treatment of cells with peptide showed decrease in EU signal in the nucleus (b) Quantification of transcription inhibition in MMGP1-treated C. albicans by flow cytometry (X 2 - C. albicans cells showing TMR-A fluorescence i.e cells that are transcriptionally active).

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Inhibition, Staining, Fluorescence, Flow Cytometry

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) ROS induction in C. albicans cells treated with MMGP1. 1-C . albicans cells without MMGP1 (negative control panel); 2-C . albicans cells treated with MMGP1 for 6 h (Test panel); 3- C. albicans cells treated with H 2 O 2 for 6 h (b) Time-scale measurement of intracellular ROS in MMGP1 treated C. albicans (0.57 µM) by flow cytometry. The fluorescence obtained with the cells treated with 1 mM of H 2 O 2 serves as positive control and the cells without peptide serves as negative control.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Negative Control, Flow Cytometry, Fluorescence, Positive Control

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Time-dependent measurement of protein carbonyls in MMGP1 treated C. albicans cells by DNPH assay. (b) Time-dependent measurement of TBARS production in MMGP1 treated C. albicans cells by TBA assay.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques:

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Measurement of mitochondrial membrane potential in MMGP1 treated C. albicans cells by flow cytometry (b) Measurement of inner mitochondrial membrane depolarization by MMGP1 in C. albicans cells. 1-mitochondria of C. albicans cells without treatment; 3-mitochondria of C. albicans cells treated with 1 mM H 2 O 2 ; 2-mitochondria of C. albicans cells treated with MMGP1 for 24 h.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: Membrane, Flow Cytometry

Journal: PLoS ONE

Article Title: Mechanisms of the Antifungal Action of Marine Metagenome-Derived Peptide, MMGP1, against Candida albicans

doi: 10.1371/journal.pone.0069316

Figure Lengend Snippet: (a) Confocal micrographs of TUNEL stained C. albicans cells treated with MMGP1 for 0, 6, 12, and 24 h (20 x magnification). The images are overlay of TUNEL (green fluorescence), Hoechst 33342 (blue) and bright field micrographs of C. albicans cells (b) Flow cytometry analysis of TUNEL stained MMGP1-treated C. albicans cells at different time intervals (0, 6, 12, and 24 h). The cells treated with H 2 O 2 was used a TUNEL positive cells.

Article Snippet: MMGP1 peptide was synthesized with >98% purity employing solid phase methods using N -(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry (Genscript Corporation, Piscataway, NJ, USA).

Techniques: TUNEL Assay, Staining, Fluorescence, Flow Cytometry