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. 2022 Oct 20;13:6239. doi: 10.1038/s41467-022-33862-0

Fig. 2. EVs secreted by MMA-treated fibroblasts increase tumor aggressiveness.

Fig. 2

a Immunoblots of A549 and A375 tumor cells after 5-day treatment by conditioned media from vehicle or MMA-treated MRC-5 (for A549) or BJ (for A375) fibroblasts. b A mixture of vehicle- or MMA-treated MRC5 with A549 cells were injected subcutaneously. The primary tumor and metastasis formation was measured after 6 weeks (n = 9 mice, mean ± SEM, two-sided unpaired t-test). c Experimental scheme. d–h Pro-aggressive properties in A549 and A375 tumor cells treated with EVsveh-MRC5/BJ or EVsMMA-MRC5/BJ from MRC-5 (for A549) or BJ fibroblasts (for A375), evaluated by immunoblots measuring EMT marker expression (d), drug resistance assays using carboplatin and paclitaxel for A549 cells, and vemurafenib and AZD6244 for A375 cells (e; n = 3 independent experiments, mean ± SEM, two-way ANOVA), colony formation assays for 3 weeks (f; n = 4 independent experiments, mean ± SEM, two-sided paired t-test), transwell invasion and migration assays (g; red scale bar indicates 100μM, n = 3 independent experiments, mean ± SEM, two-sided paired t-test) and measurement of primary tumor and metastasis formation 5 weeks after subcutaneous injection into mice (h; n = 10 mice for A549 EVsveh-MRC5 group, n = 9 mice for A549 EVsMMA-MRC5 group, n = 8 mice for A375 EVsveh-BJ group, n = 9 mice for A375 EVsMMA-BJ group, mean ± SEM, two-sided unpaired t-test).