1. Volume 25, Issue 7, Pages 1596-1605 (July 2017)
Circulating Lipoproteins: A Trojan Horse Guiding Squalenoylated Drugs to LDL- Accumulating Cancer Cells 
Dunja Sobot, Simona Mura, Marie Rouquette, Branko Vukosavljevic, Fanny Cayre, Eric Buchy, Grégory Pieters, Sébastien Garcia- Argote, Maike Windbergs, Didier Desmaële, Patrick Couvreur 
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. Molecular Therapy 2017 25, 1596-1605DOI: (10.1016/j.ymthe.2017.05.016)
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

3. Figure 1
Structures of the Bioconjugates and Physico-chemical Properties of the NPs Used in This Study
(A) Chemical structures of SQGem, 2H-SQGem, and 3H-SQGem bioconjugates, and schematic representation of the corresponding unlabeled, deuterated, and tritiated NPs prepared according to the nanoprecipitation technique. Color changes as compared with the unlabeled bioconjugate highlight chemical modification on the SQ (green) or the Gem (yellow) moiety. (B) Mean diameter and polydispersity index (PDI) of SQGem, 2H-SQGem, and 3H-SQGem NPs.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

4. Figure 2 SQGem NPs and Gem Cytotoxicity
(A–D) Cell viability of (A) A-549, (B) SK-OV-3, (C) MCF-7, and (D) MDA-MB-231 cells treated with increasing concentrations of Gem as free drug or in the form of SQGem NPs for 72 hr at 37°C. Values represent mean ± SD.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

5. Figure 3 Raman Microscopy Spectra
(A) Single Raman spectra of nucleus, cytoplasm, and intracellular lipid droplets in MDA-MB-231 cells. (B) Raman spectra of lipid droplets, SQGem NPs, and 2H-SQGem NPs. The region of interest is highlighted.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

6. Figure 4
Confocal Raman Images of MDA-MB-231 and MCF-7 Breast Cancer Cell Lines Showing a Comparison of 2H-SQGem NPs Uptake
(A and B) Representative images of non-treated MDA-MB-231 (A) and MCF-7 (B) cells (control). (C and D) MDA-MB-231 (C) and MCF-7 (D) cells incubated with NPs (77 μM) for 2 hr at 37°C. False-color Raman images were generated based on different scattering patterns of different cellular compartments. False colors visualize nucleus in dark blue, cytoplasm in white, lipid vesicles in cyan, and 2H-SQGem in pink. Scale bars, 10 μm.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

7. Figure 5 3H-SQGem NPs Uptake in MDA-MB-231, MCF-7, and MCR-5 Cells
Comparison of 3H-SQGem NPs uptake after 30 min, 2 hr, 4 hr, and 6 hr incubation at 37°C. SQGem concentration: 10 μM. Results are expressed as nanomoles of Gem per million of cells. Data represent mean ± SEM (*p < 0.05; **p < 0.01; ***p < 0.001).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

8. Figure 6 Uptake of 3H-SQGem NPs and 3H-Gem in the MDA-MB-231 Cell Line
(A) Comparison of 3H-SQGem NPs (10 μM) uptake at 37°C and 4°C. (B) Comparison of 3H-SQGem NPs (10 μM) and 3H-Gem (10 μM) uptake at 37°C. Results are expressed as nanomoles of Gem per million cells. Bars represent mean ± SEM (**p < 0.01; ***p < 0.001; n = 3).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

9. Figure 7
Cell Uptake of 3H-SQGem NPs and 3H-Gem as a Function of LDLR Expression and Availability
(A–C) MDA-MB-231 cells were cultured for 24 hr in medium supplemented with FBS, LPDS, or an excess of LDLs in LPDS-supplemented medium (LPDS + LDL) and then incubated with 3H-SQGem NPs (10 μM) or 3H-Gem (10 μM) for (A) 30 min, (B) 2 hr, or (C) 8 hr. Before addition to cells, 3H-SQGem NPs and 3H-Gem were diluted with FBS-supplemented medium (solid purple or blue bars, respectively) or LPDS-supplemented medium (square pattern purple or blue bars, respectively) and pre-incubated in these media for 30 min at 37°C. (D) MDA-MB-231 cells were cultured for 24 hr in medium supplemented with FBS or LPDS and then incubated with 3H-SQGem NPs (10 μM) (striped purple bars) or 3H-Gem (10 μM) (striped blue bars) for 30 min. Before addition to cells, 3H-SQGem NPs and 3H-Gem were diluted with pure medium and pre-incubated for 30 min at 37°C. Results are expressed as nanomoles of Gem per million cells. Bars represent mean ± SEM.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions

10. Figure 8 Tumor Growth Inhibition in MDA-MB-231 Tumor-Bearing Mice
All groups received four intravenous injections on days 0, 4, 8, and 14 in the lateral tail vein of: (1) SQGem NPs (10 mg/kg equivalent Gem), (2) Gem (10 mg/kg), or (3) dextrose 5% (control [Ctrl]). Tumor volume was regularly measured during the experimental period. The values represent mean ± SEM (n = 6). After 25 days, statistical analysis of tumor volume ratios showed superior antitumor efficacy of SQGem NPs compared with the other treatments (**p < 0.01; ***p < 0.001). Arrows point to treatment days.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2017 The American Society of Gene and Cell Therapy Terms and Conditions