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

Fig. 5. TGF-β and NF-κB mediated activation of fibroblasts and EV-associated IL6 secretion occurs downstream of ROS generation.

Fig. 5

a (left) ROS levels and (right) MDA levels in MRC-5 fibroblasts after 1 mM MMA treatment (n = 4 independent experiments for ROS measurement, mean ± SEM, one-way ANOVA; n = 3 independent experiments for MDA measurement, two-sided paired t-test). b ROS levels in MRC-5 fibroblasts after MMA treatment alone or in combination with NAC or SkQ1(n = 4 independent experiments, mean ± SEM, two-sided paired t-test). c, d Immunoblots of MRC-5 fibroblasts treated with MMA alone or in combination with NAC or SkQ1 for 6 h (c) or 5 days (d). e Immunoblots showing IL-6 amount in EVs from MRC-5 fibroblasts after MMA treatment alone or in combination with NAC or SkQ1. f Immunoblots measuring signaling activation in A549 cells treated for 3 h with EVsveh-MRC5 or EVsMMA-MRC5from MRC-5 fibroblasts treated with MMA alone or in combination with NAC or SkQ1. g, h Pro-aggressive traits of A549 cells treated for 5 days with EVsveh-MRC5 or EVsMMA-MRC5 from MRC-5 fibroblasts treated with MMA alone or in combination with NAC or SkQ1, evaluated by immunoblots measuring EMT marker expression (g) and carboplatin resistance assay (h); n = 3 independent experiments, mean ± SEM, two-way ANOVA). i Primary tumor and metastasis formation in mice 4 weeks after subcutaneous injection of A549 cells treated with EVs from MRC-5 fibroblasts treated with vehicle, MMA, or MMA and NAC (n = 10 mice for EVsveh-MRC5 and EVsMMA-MRC5+NAC groups, n = 9 mice for EVsMMA-MRC5 group, mean ± SEM, two-sided unpaired t-test). Data are partially previously represented in Fig. 2g.