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

Fig. 4. MMA promotion of the CAF phenotype and EV-associated IL6 secretion occurs through TGF-β and NF-κB signaling.

Fig. 4

a Pathway enrichment analysis on RNA-seq data from MRC-5 cells treated with vehicle or MMA for 5 days. Genes with expression differences greater than two-fold and p < 0.001 were counted (n = 3 independent experiments, two-sided paired t-test). b TGFβ signaling leads to downstream NF-κB activation. c Immunoblots measuring signal activation over time in MRC-5 fibroblasts treated with 1 mM MMA. d Immunoblots measuring signaling activation in MRC-5 fibroblasts treated with MMA alone or in combination with IKK inhibitor IKK16, TGFβR inhibitor SB431542, or TAK1 inhibitor TAKinib for 6 h. e Immunoblots of CAF markers in MRC-5 lysates treated with MMA in combination with IKK inhibitor IKK16, TGFβR inhibitor SB431542, or TAK1 inhibitor TAKinib for 5 days. f Immunoblots showing IL-6 amount in EVs from MRC-5 fibroblasts treated with vehicle or MMA or MMA alone or in combination with IKK inhibitor IKK16, TGFβR inhibitor SB431542, or TAK1 inhibitor TAKinib. g Immunoblots measuring signaling activation in A549 cells treated with EVs from MRC-5 fibroblasts treated with MMA alone or in combination with IKK inhibitor IKK16, TGFβR inhibitor SB431542, or TAK1 inhibitor TAKinib for 3 h. h, i Pro-aggressive properties of A549 cells treated with EVs from MRC-5 fibroblasts treated with vehicle, MMA alone, or MMA in combination with IKK inhibitor IKK16, TGFβR inhibitor SB431542, or TAK1 inhibitor TAKinib, evaluated by immunoblots measuring EMT marker expression (h) and carboplatin resistance assay (i; n = 3 independent experiments, mean ± SEM, two-way ANOVA).