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. 2024 Feb 16;146(8):5118–5127. doi: 10.1021/jacs.3c08000

Figure 4.

Figure 4

Delivery of ALP-functionalized lipoproplexes and enzymatic activity assay in the cells. (A) Preparation of ALP-loaded lipoproplexes (ALP-LPX). (B) Schematic representation of the experimental procedure for ALP delivery and measurement of its activity inside the cells. (C) Flow cytometry distribution of hydrolyzed DDAO substrate inside B16 cells treated with ALP-LPX and in control samples (λEx = 640 nm; λEm = 661/19 nm). The distribution for ALP alone control is not visible, as it has practically the same shape as the one for ALP+liposomes control. (D) Normalized intracellular fluorescence intensities of hydrolyzed DDAO (mean ± s.d.) detected by fluorescence microscopy. White bar shows intrinsic ALP activity (normalized to 100%) for cells incubated with the phosphate buffer (in the absence of ALP). Black circles show the data points. (n = 3 independent experiments, Kruskal–Wallis Test and Dunn’s multiple comparison test, p = 0.0412 ALP-LPX vs ALP+liposomes and p = 0.0689 for ALP-LPX vs ALP alone). (E) Example of combined transmission and fluorescence microscopy images of hydrolyzed DDAO substrate after treatment of B16 cells with enzyme alone (ALP) or enzyme loaded into DNA nanogels (ALP-NGs) in the presence and absence of liposomes.