Journal: PLoS ONE

Article Title: COPA and SLC4A4 are Required for Cellular Entry of Arginine-Rich Peptides

doi: 10.1371/journal.pone.0086639

Figure Lengend Snippet: Analysis of FITC-9R uptake in HeLa cells transfected with siRNAs against COPA, SLC4A4, ATP8B3, and CX40.1. (A) FITC-9R uptake of HeLa cells transfected with 4 individual siRNAs that were selected by primary screening. Scale bars = 10 µm. (B) Fluorescence intensity was measured by MetaMorph. Error bars represent SD from three independent experiments.

Article Snippet: The fluorescence intensity of HeLa cells was measured by MetaMorph, which showed that COPA siRNA1, COPA siRNA2, SLC4A4 siRNA1, and SLC4A4 siRNA2 reduced the internalization of FITC-9R compared with scrambled siRNA ( ).

Techniques: Transfection, Fluorescence

Journal: PLoS ONE

Article Title: COPA and SLC4A4 are Required for Cellular Entry of Arginine-Rich Peptides

doi: 10.1371/journal.pone.0086639

Figure Lengend Snippet: (A) RNA interference for COPA and SLC4A4. Interference of each gene was executed by two distinct gene-specific siRNAs. HeLa cells were cultured with each siRNA for 72 h. FITC-9R was added 1 h at 37°C before observation, and cells were observed with confocal microscopy. Scale bars = 10 µm. (B) Fluorescence intensity was measured by MetaMorph. Error bars represent SD from three independent experiments.

Article Snippet: The fluorescence intensity of HeLa cells was measured by MetaMorph, which showed that COPA siRNA1, COPA siRNA2, SLC4A4 siRNA1, and SLC4A4 siRNA2 reduced the internalization of FITC-9R compared with scrambled siRNA ( ).

Techniques: Cell Culture, Confocal Microscopy, Fluorescence

Journal: PLoS ONE

Article Title: COPA and SLC4A4 are Required for Cellular Entry of Arginine-Rich Peptides

doi: 10.1371/journal.pone.0086639

Figure Lengend Snippet: (A) HeLa cells were transfected with COPA and/or SLC4A4 siRNAs and cultured for 72 h. Then FITC-9R was added to cells and incubated for 1 h at 37°C. Cells were observed with confocal microscopy. Scale bars = 10 µm. (B) Fluorescence intensity was measured by MetaMorph. Error bars represent SD from three independent experiments.

Article Snippet: The fluorescence intensity of HeLa cells was measured by MetaMorph, which showed that COPA siRNA1, COPA siRNA2, SLC4A4 siRNA1, and SLC4A4 siRNA2 reduced the internalization of FITC-9R compared with scrambled siRNA ( ).

Techniques: Transfection, Cell Culture, Incubation, Confocal Microscopy, Fluorescence

Journal: PLoS ONE

Article Title: COPA and SLC4A4 are Required for Cellular Entry of Arginine-Rich Peptides

doi: 10.1371/journal.pone.0086639

Figure Lengend Snippet: (A) Confocal images of EGFP-COPA and EGFP-SLC4A4 localization in HeLa cells. Scale bars, 10 µm. Right Panel, high magnification merge images. Scale bars = 2 µm. (B) Confocal microscopy images of double fluorescence imaging show the co-localization of FITC-9R with COPA or SLC4A4 in HeLa cells. HeLa cells identified by DIC (differential interference contrast), and signaling with FITC-9R (9R) (green) were also positive for COPA (red), and co-localization was evident when images were merged (yellow). Similarly, the expression of SLC4A4 co-localized with FITC-9R is shown. pDsRed empty vector was used as a negative control and did not show co-localization with FITC-9R. Scale bars, 20 µm. (C) High magnification images from Fig. 5A. Scale bars = 10 µm.

Article Snippet: The fluorescence intensity of HeLa cells was measured by MetaMorph, which showed that COPA siRNA1, COPA siRNA2, SLC4A4 siRNA1, and SLC4A4 siRNA2 reduced the internalization of FITC-9R compared with scrambled siRNA ( ).

Techniques: Confocal Microscopy, Fluorescence, Imaging, Expressing, Plasmid Preparation, Negative Control

Journal: PLoS ONE

Article Title: COPA and SLC4A4 are Required for Cellular Entry of Arginine-Rich Peptides

doi: 10.1371/journal.pone.0086639

Figure Lengend Snippet: HeLa cells were transfected COPA or SLC4A4 siRNAs, and after 72-9R or FITC-TAT were added and incubated for 1 h at 37°C. Cells were observed with confocal microscopy (A, C). Scale bars = 10 µm. Fluorescence intensity of FITC-9R and FITC-TAT were measured by MetaMorph (B, D). Error bars represent SD from three independent experiments.

Article Snippet: The fluorescence intensity of HeLa cells was measured by MetaMorph, which showed that COPA siRNA1, COPA siRNA2, SLC4A4 siRNA1, and SLC4A4 siRNA2 reduced the internalization of FITC-9R compared with scrambled siRNA ( ).

Techniques: Transfection, Incubation, Confocal Microscopy, Fluorescence