Journal: ACS nano

Article Title: Overcoming Tamoxifen Resistance of Human Breast Cancer by Targeted Gene Silencing Using Multifunctional pRNA Nanoparticles

doi: 10.1021/acsnano.6b05910

Figure Lengend Snippet: Construction and characterization of pRNA–HER2apt–siMED1 nanoparticles. (A) Scheme of the pRNA–HER2apt–siMED1 (p-HER2-siMED1) structure. (B) p1 and p2 strands of pRNA–HER2apt–siMED1were transcribed using an in vitro RNA transcription system and separated in 8% denatured PAGE gel. (C) pRNA–HER2apt–siMED1 nanoparticles were generated by annealing equal molar of strands p1 and p2 and subjected to 8% native PAGE gel electrophoresis. (D) DLS assay of hydrodynamic size of pRNA–HER2apt–siMED1 nanoparticle. (E) Tm value of pRNA–HER2apt–siMED1 nanoparticle determined by TGGE assay. (F) Atomic force microscopy (AFM) images of pRNA–HER2apt–siMED1 nanoparticles. (G) Stability of control unmodified and 2′-F-modified pRNA nanoparticles was examined by 8% native PAGE gel electrophoresis after RNase A, 10% FBS-supplemented DMEM medium, and 8 M urea treatments for the indicated time at 37 °C.

Article Snippet: We thank Drs. A. Lushnikov and A. Krasnoslobodtsev of the Nanoimaging Core Facility at the University of Nebraska Medical Center for assisting us with atomic force microscopy imaging and B. Ehmer and G. Doerman (University of Cincinnati) for technical and editorial support, respectively.

Techniques: In Vitro, Generated, Clear Native PAGE, Nucleic Acid Electrophoresis, Dynamic Light Scattering Assay, Microscopy, Control, Modification

Journal: ACS nano

Article Title: Overcoming Tamoxifen Resistance of Human Breast Cancer by Targeted Gene Silencing Using Multifunctional pRNA Nanoparticles

doi: 10.1021/acsnano.6b05910

Figure Lengend Snippet: pRNA–HER2apt–siMED1 nanoparticles specifically targeted BT474 cells in vitro and in vivo. (A) Confocal microscopy analyses of the internalization of AF647-labeled control and pRNA–HER2apt–siMED1 nanoparticles by BT474 cells. Scale bar: 10 μm. (B) Flow cytometry assays of the cellular uptake of AF647-labeled control and pRNA–HER2apt–siMED1 nanoparticles by BT474 cells. (C) IVIS Lumina live imaging of BT474 orthotopic xenograft mice 24 h after i.v. injection of indicated AF647-labeled pRNA nanoparticles (10 mg/kg). (D) Major organs and tumors of above mice were excised and imaged for AF647 fluorescence. (E) Frozen tumor sections were examined for localization of AF647-labeled pRNA nanoparticles (red) using confocal microscopy. The blood vessels were stained with anti-CD31 primary antibody and Alexa488-conjugated secondary antibody (green). The nuclei were stained with DAPI (blue). Scale bar: 50 μm. (F) The fluorescence intensity of AF647-labeled pRNA nanoparticles (red) in frozen tumor sections was quantified with Image-pro Plus software.

Article Snippet: We thank Drs. A. Lushnikov and A. Krasnoslobodtsev of the Nanoimaging Core Facility at the University of Nebraska Medical Center for assisting us with atomic force microscopy imaging and B. Ehmer and G. Doerman (University of Cincinnati) for technical and editorial support, respectively.

Techniques: In Vitro, In Vivo, Confocal Microscopy, Labeling, Control, Flow Cytometry, Imaging, Injection, Fluorescence, Staining, Software