Journal: International Journal of Nanomedicine

Article Title: Self-assembled nanoparticles based on the c(RGDfk) peptide for the delivery of siRNA targeting the VEGFR2 gene for tumor therapy

doi: 10.2147/ijn.s63717

Figure Lengend Snippet: Figure 4 Confocal laser scanning microscopy images of different cell lines and different transfection time in HUVECs. Notes: (A) Different cell lines after incubation with the RPM/siRNA complexes for 6 hours. (B) The cellular uptake and localization of RPM/siRNA complexes in HUVECs at 1 or 6 hours after transfection. Cell nuclei were counterstained with DAPI (blue) and siRNA was labeled with Cy5 (red). Scale bar is 20 μm. Abbreviations: Cy5, indodicarbocyanine-5; DAPI, 4′,6-diamidino-2-phenylindole; HUVEC, human umbilical vein endothelial cell; RPM, cyclo(Arg-Gly-Asp-d-Phe-Lys)-8- amino-3,6-dioxaoctanoic acid-β-maleimidopropionic acid; siRNA, small interfering RNA.

Article Snippet: Samples were collected at different time points, and electrophoresed in a 2% agarose gel. cell culture Human umbilical vein endothelial cells (HUVECs) (American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in DMEM supplemented with 10% FBS and a 1% antibiotic-antimycotic solution.

Techniques: Confocal Laser Scanning Microscopy, Transfection, Incubation, Labeling, Small Interfering RNA

Journal: International Journal of Nanomedicine

Article Title: Self-assembled nanoparticles based on the c(RGDfk) peptide for the delivery of siRNA targeting the VEGFR2 gene for tumor therapy

doi: 10.2147/ijn.s63717

Figure Lengend Snippet: Figure 3 Confocal laser scanning microscopy images of HUVECs that were transfected with different RPM/siRNA complexes. Notes: (A) Confocal laser scanning microscopy images of HUVECs after 6 hours’ incubation of RPM/siRNA complexes which contain different volume ratio between RPM and siRNA. (B) High-definition DIC images of HUVECs when the volume ratio between RPM and siRNA is 1.5:1. Cell nuclei were counterstained with DAPI (blue) and siRNA was labeled with Cy5 (red). Scale bar is 20 μm. Abbreviations: Cy5, indodicarbocyanine-5; DAPI, 4′,6-diamidino-2-phenylindole; DIC, differential interference contrast; HUVECs, human umbilical vein endothelial cells; RPM, cyclo(Arg-Gly-Asp-d-Phe-Lys)-8-amino-3,6-dioxaoctanoic acid-β-maleimidopropionic acid; siRNA, small interfering RNA.

Article Snippet: Samples were collected at different time points, and electrophoresed in a 2% agarose gel. cell culture Human umbilical vein endothelial cells (HUVECs) (American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in DMEM supplemented with 10% FBS and a 1% antibiotic-antimycotic solution.

Techniques: Confocal Laser Scanning Microscopy, Transfection, Incubation, Labeling, Small Interfering RNA

Journal: International Journal of Nanomedicine

Article Title: Self-assembled nanoparticles based on the c(RGDfk) peptide for the delivery of siRNA targeting the VEGFR2 gene for tumor therapy

doi: 10.2147/ijn.s63717

Figure Lengend Snippet: Figure 6 Cytotoxicity and gene silencing in vitro. Notes: (A) Cell viability by CCK-8 assay (Beyotime Institute of Biotechnology, Haimen, People’s Republic of China). (B) RT-qPCR analysis of VEGFR2 mRNA levels expressed in HUVECs after a 48-hour transfection (100 nM siRNA:siVEGFR2). All qPCR experiments were done in triplicate and VEGFR2 mRNA expression was normalized to the expression of GAPDH. (C) Western blot analysis of VEGFR2 protein and GAPDH protein expressed in HUVECs after a 72-hour transfection (100 nM siRNA:VEGFR2). Three independent experiments were performed. (D) Quantitative analysis of VEGFR2 protein expression levels. The expression of targeted protein was relative to the expression of GAPDH protein. *P0.05, **P0.01, compared with untreated; #P0.05, ##P0.01, compared with RPM; &&P0.01, compared with RPM/control siRNA;

Article Snippet: Samples were collected at different time points, and electrophoresed in a 2% agarose gel. cell culture Human umbilical vein endothelial cells (HUVECs) (American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in DMEM supplemented with 10% FBS and a 1% antibiotic-antimycotic solution.

Techniques: In Vitro, CCK-8 Assay, Quantitative RT-PCR, Transfection, Expressing, Western Blot, Control

Journal: International Journal of Nanomedicine

Article Title: Self-assembled nanoparticles based on the c(RGDfk) peptide for the delivery of siRNA targeting the VEGFR2 gene for tumor therapy

doi: 10.2147/ijn.s63717

Figure Lengend Snippet: Figure 5 Confocal laser scanning microscopy images of HUVECs that were treated with different ways. Notes: The HUVECs were incubated with anti-integrin αvβ3 antibody for 30 minutes prior to transfection with RPM/siRNA for 6 hours (A) or incubated with the RAPM/ siRNA complexes for 6 hours (B). Cell nuclei were counterstained with DAPI (blue) and siRNA was labeled with Cy5 (red). Scale bar is 20 μm. Abbreviations: c(RADfk), cyclo(Arg-Ala-Asp-d-Phe-Lys); c(RGDfk), cyclo(Arg-Gly-Asp-d-Phe-Lys); Cy5, indodicarbocyanine-5; DAPI, 4′,6-diamidino-2-phenylindole; HUVEC, human umbilical vein endothelial cell; MAL, β-maleimidopropionic acid; PEG, 8-amino-3,6-dioxaoctanoic acid; RAPM, c(RADfk)-PEG-MAL; RPM, c(RGDfk)-PEG- MAL; siRNA, small interfering RNA.

Article Snippet: Samples were collected at different time points, and electrophoresed in a 2% agarose gel. cell culture Human umbilical vein endothelial cells (HUVECs) (American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in DMEM supplemented with 10% FBS and a 1% antibiotic-antimycotic solution.

Techniques: Confocal Laser Scanning Microscopy, Incubation, Transfection, Labeling, Small Interfering RNA