Fig. 1.

Preparation, characterization, and targeting of HELA-Exos. A and B Relative abundance of α-LA mRNA in different cell lines (the human normal breast epithelial cell line MCF-10A, breast adenocarcinoma cell line MCF7, breast cancer cell line MDA-MB-435S, breast cancer cell line SKBR3, TNBC cell line MDA-MB-231, gastric adenocarcinoma cell line BGC-823, transitional cell carcinoma cell line T24, alveolar basal epithelial adenocarcinoma cell line A549, and leukemic cell line THP-1) and infected MDA-MB-231 cells by qPCR; data were normalized to GAPDH. The data are presented as the mean ± SD; n = 6. A t test was performed for statistical analysis (****: P < 0.0001). C Western blot analysis for detecting the expression of α-LA protein in cells as in A. β-Actin was used as a loading control. D FITC-labeled ELANE and CX-Rhodamine-labeled Hiltonol were electroporated with exosomes at 400 V 150 µF in the volume of buffer shown on the x-axis, and ELANE and Hiltonol loading were determined based on the fluorescent standard curve. The data are presented as the mean ± SD; n = 3. E The loading of ELANE and Hiltonol in exosomes was characterized by nanoscale flow cytometry. F Transmission electron micrograph of LA-Exos and HELA-Exos. Scale bar, 100 nm. G Size distribution of LA-Exos and HELA-Exos measured by a particle sizer. H Western blot analysis for detecting the expression of the α-LA protein and exosomal markers in Texs, LA-Exos, HLA-Exos, ELA-Exos, and HELA-Exos. I Diameter change of HELA-Exos from day 1 to day 7. The data are presented as the mean ± SD; n = 3. J Flow cytometry analysis of the cellular uptake of MDA-MB-231 cells or PBMCs after incubation with Texs-DiI or HELA-Exos-DiI for 2 h in the coculture model