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. 2021 Jun 21;118(25):e2023537118. doi: 10.1073/pnas.2023537118

Fig. 6.

Fig. 6.

TGF-β–dependent activation of the miR-218-5p ceRNA network. (A) qPCR analysis of TGF-β family members and TGF-β receptors. Values were normalized to H2122-P. (B) TGF-β1 ELISA assay on CM samples. (C) WB analysis of p-Smad3 (Ser423/425) and total Smad2/3 in cell lysates. α-Tubulin served as loading control. (D) qPCR analysis of TGF-β downstream targets (CTGF, PAI-1, and LEF1) and predicted miR-218-5p targets (LEF1, OC2, N-cad, and PI4KIIα) in H2122-P cells that had been treated for 5 d with DMSO or 10 ng/mL TGF-β1. Values were normalized to DMSO control. (E) WB analysis of cells in D. β-Actin served as loading control. (F) WB analysis of H2122-BT cells treated for 72 h with DMSO or 5 µM ALK5i. (G and H) WB analysis of H2122-P cells that were treated for 72 h with 10 ng/mL TGF-β1 and then transfected with miR-218-5p mimics (G) or siRNAs against OC2 or N-cad (H). Values are shown as mean ± SEM of triplicate biological replicates. P values: one-way ANOVA in A and B and two-sided t test in D. *P < 0.05, **P < 0.01, and ***P < 0.001.