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. 2023 Apr 4;15(7):2138. doi: 10.3390/cancers15072138

Figure 5.

Figure 5

Characterization of SKBR3 exosomes and their effect in trastuzumab response. (A) Western blot was performed for measuring the presence of TSG-101 and CD9 typical from exosomes, and the absence of calnexin as a cellular control. (B) Representative image of exosomes observed via TEM (microscope JEOL1010, 60,000× Magnification) (scale bar: 50 nm). (C) Analysis via RT-qPCR of miR-146a-5p expression in exosomes from SKBR3 (exo), exosomes from SKBR3r (exoR) line, and from SKBR3 and SKBR3r cell lines. miR-16a was used as endogenous miRNA. (D) Percentage of fluorescent cells detected via flow cytometry after exosome internalization assay with DMEM F-12 alone or exoR containing (E), and mean of fluorescence detected in cells treated or not with trastuzumab. (F) Viability assay of the SKBR3 cell line co-cultured with 50% of CM from SKBR3r or exoR, treated with or without 15 µg/mL trastuzumab for 7 days. (G) Viability assay of the SKBR3 cell line co-cultured with exoR after transfection with miR-146a-5p inhibitor with or without 15 µg/mL trastuzumab for 7 days. Each experiment was performed in technical and biological triplicate. * p-value < 0.05; *** p-value < 0.001. The uncropped blots are shown in Figure S5.