Fig. 4: Evaluation of gene editing efficiency and therapeutic effect in vitro models of RDN@PL.
From: Reactive oxygen species responsive nanomotors for gene edited metabolic disruption and immunotherapy
a The images of GFP green fluorescent protein-expressing of B16F10 cells after transfection by different particles (Scale bar: 200 μm), and the corresponding (b) statistical analysis of fluorescence intensity (n  =  3 independent experiments). c T7E1 indel analysis of B16F10 cells after Non-RDN@PL and RDN@PL mediated transfection of CRISPR/Cas9 plasmid targeting LDHA locus. The experiment was repeated independently three times with similar results, and a representative result is shown. d Schematic of the Transwell model in vitro to mimic the tumor vascular barrier, with Cy5.5-labeled particles added in the inner chamber. e The statistical analysis of fluorescence intensity in outer nest after particles addition for 12 h (n  =  3 independent experiments). f The statistical analysis of HUVECs’ cellular activity in inner chamber and B16F10 cells in outer nest after 24 h incubation with particles (n  =  3 independent experiments). g Illustration of the 3D MTSs treatment by RDN@PL. h 3D view and Z-stack CLSM images of Cy5.5-labeled particles distributed in MTSs (Scale bar: 200 μm), and corresponding (i) Fluorescence distribution curves of Z = 80 μm. j The fluorescence images of MTSs transfected with different particles (Scale bar of merge images: 200 μm) The experiment was repeated independently three times with similar results, and a representative result is shown. k The frequency of indel mutation determined by T7E1 assay of MTSs with treatment. The experiment was repeated independently three times with similar results, and a representative result is shown.
