Fig. 2: The preparation and characterization of sgPik3cg-DHP/DGA-NVs.
a A total of 1 × 109 colony forming unit (CFU) E.coli derived protoplasts were added with indicated amounts of RhB-DHP and Cy5.5-DGA for physical extrusion. The decoration efficiency and quantitative analysis of RhB-DHP and Cy5.5-DGA for nanovesicles were determined through flow cytometry analysis. n = 3 biologically independent samples. b Representative TEM images illustrate sgPik3cg-NVs, both with and without DHP/DGA decoration. Scale bar, 200 nm. c The diameters of sgPik3cg-NVs and sgPik3cg-DHP/DGA-NVs were analyzed using Nanosight. For (b, c), experiments were independently conducted three times with similar results. d The ratio of RhB-positive NVs was quantified through flow cytometry analysis after treating 1 × 1010 CFSE-labeled NVs with RhB-DHP and DGA in 100 μL of pH 6.5 PBS at indicated time points (0 h and 24 h). e Quantitative analysis of sgPik3cg-RhB-DHP/Cy5.5-DGA-NVs treated with different pH values of PBS at indicated time points. n = 3 biologically independent samples for (d, e). Statistical analysis was performed using two-way ANOVA with Bonferroni’s multiple comparison test. f Direct injection of 50 μL PBS containing 5 × 109 CFSE-labeled sgPik3cg-RhB-DHP/DGA-NVs into 4T1 tumor tissues and tumor-adjacent tissues. Corresponding samples were harvested from tumor-bearing mice at 24 h post-injection for fluorescence photography. The representative images presented are from a sample size of n = 3 mice. Scale bar = 50 μm. g Measurement of IL-6 and TNF-α levels in 100 μL serum of 4T1 tumor-bearing mice at 2 h and 24 h after in vein injections of 100 μL PBS containing different types of NVs (dose: 1 × 1010 per mouse). n = 5 mice per group. Statistical analysis was performed using one-way ANOVA with Dunnett’s multiple comparison test. Data are represented as mean ± SD. *P < 0.05, **P < 0.01 and ***P < 0.001, ns, no significant change. The exact P-value and source data are provided as a Source Data file.
