Journal: PLoS ONE

Article Title: Identification of a Short Cell-Penetrating Peptide from Bovine Lactoferricin for Intracellular Delivery of DNA in Human A549 Cells

doi: 10.1371/journal.pone.0150439

Figure Lengend Snippet: (A) Images of the FITC-labeled CPP-mediated delivery of the pCAGGS-tdTomato plasmid DNA into cells. L6-FITC or L5a-FITC was premixed with the pCAGGS-tdTomato plasmid at an N/P ratio of 12. These CPP/DNA complexes, DNA alone and CPP alone were incubated with A549 cells for 1 h. Untreated cells and cells treated with Lipofectamine 2000/DNA complexes served as negative and positive controls, respectively. Following the 1 h incubation, cells were washed with PBS and incubated at 37°C for an additional 48 h. BFP, GFP, and RFP channels displayed the distribution of nuclei stained by Hoechst33342, CPP-FITC, and tdTomato, respectively. Cell morphologies were observed in bright-field images. All images are obtained using a Nikon Eclipse 80i fluorescent microscope at a magnification of 1,000×. (B) Expression of reporter gene delivered by CPPs. Fluorescent intensity recorded in panel A was quantified using UN-SCAN-IT software, and analyzed by ANOVA. Data are presented as mean ± SD from three independent experiments in each treatment group. Experimental groups were compared with the negative control. Significant differences from the negative control at P <0.05 (α) and P <0.01 (**, αα) are indicated. (C) Transfection efficiency of CPP-mediated delivery of gene expression. Fold increases of transfection efficiency recorded in panel A were quantified using UN-SCAN-IT software. Statistical comparisons were performed by ANOVA. Data are presented as mean ± SD from three independent experiments in each treatment group. Significant differences from the negative control at P <0.05 (α) and P <0.01 (**) are indicated.

Article Snippet: Fluorescent and bright-field images were recorded using an AE31 inverted Epi-fluorescence microscope (Motic, Causeway Bay, Hong Kong) with an IS1000 eyepiece (Tucsen, Fujian, China) or an Eclipse 80i Epi-fluorescence microscope (Nikon, Tokyo, Japan) with the NIS-elements documentation software (Nikon).

Techniques: Labeling, Plasmid Preparation, Incubation, Staining, Microscopy, Expressing, Software, Negative Control, Transfection, Gene Expression

Journal: PLoS ONE

Article Title: Bacteriophage Mediated Killing of Staphylococcus aureus In Vitro on Orthopaedic K Wires in Presence of Linezolid Prevents Implant Colonization

doi: 10.1371/journal.pone.0090411

Figure Lengend Snippet: A) Naked wire [not immersed in bacterial suspension] stained with fluorescent dye as viewed under 10x,Nikon eclipse 80i B) naked wire as seen with adhered bacteria (immersed for 6 h in bacterial susupension,10 7 CFU/ml) C) naked wire as seen with adhered bacteria (immersed for 24 h in bacterial susupension,10 7 CFU/ml) D) and E) Syto 9 stained HPMC coated wires (not immersed in bacterial suspension) showing coating on the wire F) and G) HPMC coated wires (immersed for 6 h in bacterial susupension,10 7 CFU/ml) as seen with adhered bacteria (seen as green spots) and H) HPMC coated wires as seen with adhered bacteria (immersed for 24 h in bacterial susupension,10 7 CFU/ml). (Note: Although the coating was uniform but the white arrows show slight discrepancy in the coating at some locations on wires)

Article Snippet: All the stained wires were visualized under epi-fluorescence microscope (Nikon Eclipse 80i).

Techniques: Suspension, Staining, Bacteria