Fig 1: Amphiregulin Induces Pericyte Differentiation by Releasing Bioactive TGF-ß via Integrin-aV(A and B) Primary lung pericytes were cultured in the presence or absence of 100 ng/ml Amphiregulin (A), then incubated with latent TGF-ß in the presence and absence of PLC? inhibitor U-73122 (B) and analyzed for LAP binding by flow cytometry.(C–F) After 24 h of treatment, the release of bioactive TGF-ß (upper panel), as well as their differentiation into myofibroblasts (lower panels), was determined in the presence or absence of inhibitors for the EGFR (Gefitinib) (C), PLC? inhibitor (U-73122) (D), integrin-aV (CWHM-12 and its inactive control enantiomer CWHM-96) (E), or TGF-ß-R (ALK5i) (F).(G) The induction of aSMA in treated pericytes was also evaluated at the protein level by western blot analysis.(H) Differentiation of primary lung pericytes into myofibroblasts following o/n co-culture with (left graph) or o/n exposure to supernatants derived from (right graph) alveolar macrophages isolated from infected or uninfected WT or Areg-/- mice.All data are representative of at least two independent experiments except for (B)–(D) (mean ± SEM); results for preparations from individual mice are shown as dots. See also Figure S4.