Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: A schematic diagram showing ( A ) the formation of EPSLR complexes and ( B ) the intracellular trafficking for siRNA delivery. Notes: EPSLR is associated with MCF-7/ADR cells followed by cell internalization by anti-EphA10-mediated endocytosis. Chol–SIB–PEG–Eph degraded in acidic endosome and liposome–siRNA complexes subsequently escaped from endosomes through fusion interaction. Lastly, the siRNA was released into cytoplasm and formed RISC gene expression. Abbreviations: LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody; RISC, RNA-induced silencing complex; EDCI, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydro chloride; sulfo-NHS, N-Hydroxysulfosuccinimide sodium salt; Eph, anti-EphA10 antibody; PSL, PEGylated lipsomes; EPSL, anti-EphA10 antibody coated PSL; Chol-SIB-PEG, cholesterol–Schiff base–polyethylene glycol.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: Expressing, Small Interfering RNA

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: Encapsulation efficiency and protection by EB (RiboGreen) interaction and gel retardation assay. Notes: ( A ) Gel retardation of LR, PSLR, and EPSLR at different N/P ratios; ( B ) heparin-resistant assay with different N/P ratios; ( C ) encapsulation efficiency of LR, PSLR, and EPSLR at various N/P ratios; ( D ) stability in serum of naked siRNA and liposome–siRNA. Abbreviations: LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody; EB, ethidium bromide; N/P ratios, molar ratio of DOTAP-nitrogen atoms to siRNA-phosphate.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: Electrophoretic Mobility Shift Assay, Small Interfering RNA

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: Physicochemical characteristics of complexes. Notes: The particle sizes (columns) and zeta potentials (lines) of EPSLR at different N/P ratios ( A ) and its reference formulations at an N/P ratio of 9 ( B ). Morphology of Eph-modified liposomes was characterized with TEM after staining with phosphotungstic acid ( C ). Abbreviations: LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody; TEM, transmission electron microscopy; N/P ratios, molar ratio of DOTAP-nitrogen atoms to siRNA-phosphate; Eph, anti-EphA10 antibody.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: Modification, Staining, Small Interfering RNA, Transmission Assay, Electron Microscopy

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: CLSM images of MCF-7/ADR cells incubated with naked siRNA ( A ), FAM-labeled LR ( B ), PSLR ( C ), and EPSLR ( D ). Notes: Blue and green colors indicate Hoechst 33258 and FAM-siRNA, respectively. Scale bar =10 μm. Abbreviations: CLSM, confocal laser scanning microscopy; LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

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

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: Flow cytometry measurement of intracellular uptake of FAM-siRNA-loaded complexes ( A ) and anti-EphA10 antibody competitive assay ( B ) in MCF-7/ADR cells ( P <0.01). Note: ** P <0.01. Abbreviations: LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody; Eph, anti-EphA10 antibody.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: Flow Cytometry, Small Interfering RNA

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: CLSM images of the intracellular distribution of FAM-siRNA. Notes: For each panel, images from left to right show nuclei stained by Hoechst 33258 (blue), FAM-siRNA (green), lysosome stained with LysoTracker (red) and merged images. MCF-7/ADR cells were incubated with naked siRNA ( A ), LR ( B ), PSLR ( C ), and EPSLR ( D ) for 4 hours. Scale bar =10 μm. Abbreviations: CLSM, confocal laser scanning microscopy; LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: Staining, Incubation, Confocal Laser Scanning Microscopy, Small Interfering RNA

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: In vitro cytotoxicity studies of LR, PSLR, and EPSLR. Notes: Studies of LR, PSLR, and EPSLR for 24 ( A ), 48 ( B ), and 72 hours ( C ) ( P <0.05) and Western blot. Quantification of Western blot bands using ImageJ ( D ). MDR1 silencing mediated by control, naked siRNA, LR NC , LR, PSLR, and EPSLR ( E ). Abbreviations: LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody; LR NC , Liposome–negative control siRNA complexes; N/P ratios, molar ratio of DOTAP-nitrogen atoms to siRNA-phosphate; P-gp, P-glycoprotein.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: In Vitro, Western Blot, Small Interfering RNA, Negative Control

Journal: International Journal of Nanomedicine

Article Title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA

doi: 10.2147/IJN.S107952

Figure Lengend Snippet: In vivo fluorescence images of nude mice. Notes: ( A ) Nude mice bearing MCF-7/ADR cells after tail vein administration of DIR-loaded LR, PSLR and EPSLR. ( B ) Ex vivo fluorescence images of tumors and organs collected at 24 hours postinjection of LR, PSLR, and EPSLR. ( C ) Quantification of excised organs and tumor uptake characteristics of nanocomplexes. Uptake expressed as fluorescence per mm 2 of tumor and organs. Data expressed as mean values ± SD (n=3, * P <0.05). Abbreviations: LR, liposome–siRNA complexes; siRNA, small interfering RNA; PSLR, PEGylated LR; EPSLR, PSLR-conjugated anti-EphA10 antibody; SD, standard deviation; DIR, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide.

Article Snippet: Anti-EphA10 antibody was purchased from Bioss Biotechnology Co. Ltd. (Beijing, People’s Republic of China).

Techniques: In Vivo, Fluorescence, Ex Vivo, Small Interfering RNA, Standard Deviation