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. 2017 Mar 16;7(5):1333–1345. doi: 10.7150/thno.17092

Figure 4.

Figure 4

In vitro and in vivo effects of BCR-ABL siRNA loaded IL3L-exosomes. (A) LAMA84, K562 and Imatinib resistant K562 cell growth was measured by MTT assay after 24, 48 or 72 h of treatment with Imatinib (on the right side of each histograms) or with 1 μg/ml of exosomes derived from IL3L-HEK293T containing scrambled siRNA or BCR-ABL siRNA. The values were plotted as % of growth vs Ctrl (untreated cells - dot line). Each point represents the mean ± SD of three independent experiments. Statistically significant differences were found between 1 μg/ml IL3L Exo with BCR-ABL and 1 μg/ml IL3L Exo with scramble values (Sidak test: ***, p < 0.0005). In addition, significant decrease in MTT value was observed in each cell type overtime (Sidak test: a, 48h versus 24h - p < 0.0005; b, 72h versus 48h - p < 0.0005). (B) Real-time PCR analysis was performed on CML cell lines treated for 24 or 48 h with Imatinib (on the right side of each histograms) or with 1 μg/ml of exosomes derived from IL3L-HEK293T containing scrambled siRNA or BCR-ABL siRNA. The values were plotted as fold change compared to control (untreated cells - dot line). Each point represents the mean ± SD of three independent experiments. GLM highlighted a significant influence on PCR data by the treatment effect (***, p < 0.0005), but not by experimental time effect. (C) Western blot analysis was performed on CML cell lines treated for 72h with Imatinib or with 1 μg/ml of exosomes derived from IL3L-HEK293T containing scrambled siRNA or BCR-ABL siRNA. Protein levels of Bcr-Abl were evaluated. Blots were stripped and subsequently re-probed with an antibody against Gapdh to ensure equal loading. (D) Comparison of the mean tumor volume was an index of the anti-tumor efficacy of IL3L-HEK293T derived exosomes containing BCR-ABL siRNA. Significant differences in terms of tumor volume were observed at 28 days: IL3L BCR-ABL siRNA versus Ctrl untreated (a, p < 0.0005), IL3L Exo-scrambled siRNA and Imatinib (b, p < 0.005), and versus Exo-scrambled siRNA (c, p < 0.005).