1. Volume 20, Issue 3, Pages 601-608 (March 2012)
In Vivo Delivery of Nucleic Acids via Glycopolymer Vehicles Affords Therapeutic Infarct Size Reduction In Vivo 
Michael Tranter, Yemin Liu, Suiwen He, James Gulick, Xiaoping Ren, Jeffrey Robbins, W. Keith Jones, Theresa M. Reineke 
Molecular Therapy 
Volume 20, Issue 3, Pages (March 2012)
DOI: /mt
Copyright © 2012 The American Society of Gene & Cell Therapy Terms and Conditions

2. Figure 1
Characterization of glycopolymer–ODN NF-κB decoy polyplexes. (a) The sizes of PGAA/decoy polyplexes examined by transmission electron microscopy (TEM). (b) The ability of glycopolymers to protect decoys from DNase digestion assessed by gel electrophoresis (band disappearance indicates DNA degradation). (c) Polyplex stability, as a function of increasing NaCl concentration, represented by the fraction of PicoGreen excluded from the polyplex structure (more dye intercalates into DNA upon polyplex dissociation). ODN, oligodeoxynucleotide; PGAA, poly(glycoamidoamine).
Molecular Therapy  , DOI: ( /mt )
Copyright © 2012 The American Society of Gene & Cell Therapy Terms and Conditions

3. Figure 2
Glycopolymer–ODN efficacy and toxicity in H9c2 cells. Cells were treated with increasing doses of either (a) decoy only or (b) T4–decoy polyplexes 24 hours prior to TNF-α treatment to activate NF-κB. Activation of NF-κB was assessed by EMSA. (c) IC50 doses for NF-κB decoys alone (naked decoy) and polyplexes formed with T4, M4, G4, and D4 polymers, Lipofectamine 2000, and JetPEI were calculated from sigmoidal dose-response curves fitted to EMSA results. (d) Nuclear localization of fluorescently labeled decoys in H9c2 cells were transfected alone or via T4 and the fraction of fluorescently positive nuclei were quantified at the indicated times. (e) MTT assay was used to assess cytotoxicity of delivery vectors. Error bars represent 95% confidence intervals. *P ≤ 0.05 versus all other delivery vectors (c–e). H9c2 cells were transfected with CMV-Luciferase reporter pDNA using either T4 or Lipofectamine 2000 and (f) the fraction of cell survival as well as (g) the activity of the luciferase reporter (relative to the initially measured activity 24 hours after treatment) were tracked for ten days following transfection. *P ≤ 0.05 versus baseline control. #P ≤ 0.05 versus Lipofectamine transfected group. EMSA, electrophoretic mobility shift assay; ODN, oligodeoxynucleotide.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2012 The American Society of Gene & Cell Therapy Terms and Conditions

4. Figure 3
Glycopolymer–ODN polyplex delivery and toxicity in neonatal rat ventricular myocytes. (a) Uptake of fluorescently (FITC) labeled decoy was tracked by flow cytometry. (b) Confocal microscopy of NRVMs 24 hour after transfection with FITC-labeled decoy (green). A Troponin I-specific antibody (red) was used to identify cardiomyocytes, and all cells were labeled with a nucleus-specific stain (TO-PRO-3; blue). (c) Adenylate kinase release into the culture media was assayed as a cell death marker 2, 4, 6, and 8 hours after transfection. NRVMs, neonatal rat ventricular myocytes; ODN, oligodeoxynucleotide.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2012 The American Society of Gene & Cell Therapy Terms and Conditions

5. Figure 4
Pericardial delivery of glycopolymer–ODN polyplexes in vivo efficiently silences NF-κB-dependent gene expression and reduces infarct size after I/R injury. Cardiac sections from mice 24 hours after pericardial injection of (a) Alexa488-labeled NF-κB decoy alone or (b) with T4 polymer. Quantitation of Alexa488 uptake to the myocardium by (c) mean pixel intensity and (d) percentage of fluorescent signal positive myocardium (*P ≤ 0.01). (e) Blockade of Cox-2 mRNA expression via pericardial delivery of NF-κB decoy/T4 polyplexes 24 hours prior to intraperitoneal injection of cytokine mixture. Cox-2 mRNA levels were assessed after 3 hours by quantitative real-time polymerase chain reaction. *P ≤ versus 0 µg NF-κB decoy/T4. (f) Assessment of the impact on infarct size of 10 µg ODN/T4 delivered via pericardial injection 24 hours prior to a 45-minute coronary occlusion. Results obtained from NF-κB dominant-negative (DN) mice shown for comparison. *P ≤ 0.01 vs. sham-injected C57 wild-type mice. All error bars represent SEM. I/R, ischemia/reperfusion; ODN, oligodeoxynucleotide.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2012 The American Society of Gene & Cell Therapy Terms and Conditions