Fig. 3 Characterization of LNPs and their modification with AMBPs. (a) Dynamic light scattering (DLS) of native LNPs (approximately 1.5 × 1012 LNPs/mL) or AMBP-modified LNPs (10 μM AMBP concentration, 100 μL protein per 1 mL of LNPs). (b) Cryo-TEM images of unmodified LNPs (approximately 1.5 × 1013 LNPs/mL) and (c) 1:2-LNPs (approximately 1.5 × 1013 LNPs/mL) showing successful membrane modification. Scale bars: 50 nm. (d) Phase-contrast microscopy image of giant unilamellar vesicles (GUVs) after an overnight reaction with 2 μL of 4 (10 μM). (e) Fluorescence microscopy image of GFP from (d). (f) Phase-contrast image of a single GUV modified with 4. (g) GFP fluorescence microscopy image of (f). (h) Fluorescence microscopy images (GUVs) labeled with Texas red (shown in red) and after addition of 2 μL of 5 (10 μM). (i) GFP channel of (h), shown in green. GUVs were composed of CHOL, DOPC (5% TR-DHPE only in (h) and (i)) DOPS, and DOPE in a 55:21:16:8% ratio, and lipid films were hydrated in 0.5 M sucrose and imaged in 0.5 M glucose (both in 20 mM phosphate buffer at pH 7.5). Scale bars: 50 μm.
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