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. 2020 Sep 18;218(1):e20200321. doi: 10.1084/jem.20200321

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© 2020 Derakhshan et al.

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Figure 7. — Epithelial cells induce an MMC-like phenotype through TGF-β signaling. (A) Representative flow plot of TGF-β1 expression by murine pulmonary epithelial cells (EPCAM⁺, Thy1⁻, CD90⁻, CD31⁻) in vivo from mice challenged with HDM (red) and saline (blue) relative to isotype control (gray). Results representative of three experiments with n = 2–4 mice per group in each experiment. (B) Representative flow plot of TGF-β1 expression by primary mouse tracheal epithelial cells expanded ex vivo (red) relative to isotype control (gray). Results representative of two independent experiments. (C) Representative flow plots of mMCP-1 expression by IL-3–supplemented BMMCs cultured on Matrigel alone (left) or in coculture with mouse airway epithelial cells (right) for 7 d. (D) Quantification of intracellular mMCP-1 expression by IL-3–supplemented BMMCs cultured alone or in coculture with mouse airway epithelial cells on Matrigel for 7 d. Results are from three independent experiments, with each replicate showing a unique combination of epithelial and MC biological donors. ***, P < 0.001 (t test). (E) Representative flow plots of αE integrin expression by BMMCs cultured on Matrigel alone (left) or in coculture with mouse airway epithelial cells (right) for 7 d. (F) Quantification of αE-integrin expression by BMMCs cultured on Matrigel alone or in coculture with mouse airway epithelial cells for 7 d. Results are from three independent experiments, with each replicate showing a unique biological combination of epithelial and MC cultures. ***, P < 0.001 (t test). (G) Expression of mMCP-1 and αE integrin by BMMCs cultured on Matrigel alone or in coculture with mouse airway epithelial cells for 7 d in conjunction with the indicated concentration of the TGF-β R1/R2 dual inhibitor LY2109761. Results are from three independent experiments, with each replicate showing a unique biological combination of epithelial and MC cultures. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (Dunnett’s multiple comparisons test).