1. Volume 25, Issue 7, Pages 1567-1579 (July 2017)
Dual Functional LipoMET Mediates Envelope-type Nanoparticles to Combinational Oncogene Silencing and Tumor Growth Inhibition 
Kai Shi, Yi Zhao, Lei Miao, Andrew Satterlee, Matthew Haynes, Cong Luo, Sara Musetti, Leaf Huang 
Molecular Therapy 
Volume 25, Issue 7, Pages (July 2017)
DOI: /j.ymthe
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

2. Figure 1 Schematic Representation of the Design of This Study
(A) Formation of targeted NPs. (B) Intracellular action mechanism of NPs carrying oncogene siRNA.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

3. Figure 2 Preparation and Characterization of Various NPs
(A) Typical TEM morphology and DLS size profile of the compact PH core obtained at the optimal condition. (B) Gel retardation assay of binding ability of polycation/HA to siRNA at various N/P ratios. Lane# represents double-stranded RNA ladder. (C) Typical TEM morphology of LipoEDA (a), LipoMET (b), targeted NPs (c) and LipoMET NPs (d). (D) Organization model of the two lipids with different geometric packing parameter (S) in lamellar membrane (Lα), where the cone-shaped lipid of DOPE readily adopts a hexagonal phase (HII) and MET-Chol adopts micellar phase (trans-HII).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

4. Figure 3 Confocal Evaluation of Cell Uptake and Endosomal Escape
(A–C) Intracellular trafficking of Cy3-siRNA-loaded NPs and NPs in NCI-H460 cells at various incubation times. The free Cy3-labeled siRNA was used as a control. The white arrow indicates the local close up. (D) Quantifying endosomal escape rate of Cy3-siRNA-loaded various NPs in NCI-H460 cells based on the difference of pixel intensity between the Cy3-siRNA clusters with or without co-localization with the signals of LysoTracker Green. Data are expressed as mean ± SD. *p < 0.05, **p < 0.01, n = 5.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

5. Figure 4 Intrinsic Antitumor Activity of Various Lipid Carriers
(A) Determination of effect of metformin, LipoEDA, and LipoMET on H460 cell proliferation by MTT assay (n = 6). (B) Inhibition of tumor growth in a murine model with H460 xenografts after treatment with various formulations. Arrows indicate time of injection. (C) TUNEL assay of apoptosis in tumor that induced by various formulations. (D) Quantifying TUNEL analysis of apoptosis in tumor that was induced by various formulations. (E) Expression levels of p-AMPK and p-mTOR protein in tumor tissues. Data are expressed as mean ± SD. *p < 0.05, **p < 0.01 versus control; n = 5.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

6. Figure 5
H&E Morphology Evaluation and Serum Biochemical Value Analysis
(A) H&E sections of major organs collected from various groups at the endpoint of treatments. (B) Serum biochemical value analysis. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN) and creatinine (Cr) were assayed as indicators of liver and renal functions; (C) Hematology test of whole blood. White blood cells (WBC), lymphocytes (LYMF), granulocytes (GRAN) and monocytes (MONO) were counted for the detection of myelosuppression. Data are expressed as mean ± SD. n = 5.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

7. Figure 6 Evaluation of Gene Silencing by Various NPs
(A) Luciferase gene silencing by siLuc-Lipofectamine, and in H460/Luc cells. The siCtrl complexed vehicles were used as control, respectively. (B) Inhibition of tumor growth in a murine model with NCI-H460 xenografts after treatment with various siVEGF formulations. The siCtrl functions as a negative control. Arrows indicate time of injection. (C) TUNEL assay of apoptosis in tumor tissue induced by various siVEGF formulations. (D) Quantifying TUNEL analysis of apoptosis in tumor induced by various siVEGF formulations. (E) Expression levels of VEGF protein in tumor tissues. Data are expressed as mean ± SD. *p < 0.05, **p < 0.01 versus control; n = 5.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

8. Figure 7 Tissue Distribution and Tumor Uptake of siRNA
(A) In vivo tumor uptake of Cy3-siRNA-loaded formulations in NCI-H460 xenograft tumor-bearing mice. (B) Quantification of tumor uptake of Cy3-siRNA-loaded formulations in NCI-H460 xenograft tumor-bearing mice. (C) Distribution of Cy3-siVEGF-loaded various formulations in tumor tissue and organs detected by the IVIS Imaging System. Data are expressed as mean ± SD. *p < 0.05, **p < 0.01 versus control; n = 5.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions