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. 2021 Mar 30;65(5):489–499. doi: 10.1165/rcmb.2020-0499OC

Figure 4.

Figure 4.

Noncanonical Wnt11 signaling was induced in mouse lung fibrosis. Whole-lung tissue RNA or protein lysates were collected at Day 21 after BLM or PBS treatment and were then analyzed for (A) Wnt11 mRNA by using qPCR analysis and for (B) protein by using Western blotting. (C) Total c-Jun and phosphorylated c-Jun in BLM-treated lung tissue were also assayed by using Western blotting. (D) MLFs were transfected with Cont si or Wnt11 siRNA (Wnt11 si), which was followed by TGFβ treatment. Wnt11 and α-SMA proteins were analyzed at 24, 48, and 72 hours after treatments by using Western blotting, and quantified protein levels normalized to GAPDH signals are shown in the bar graphs. (E) AECIIs and MLFs were analyzed for mRNA expression of Wnt3a, Wnt11, and Wnt5a by using qPCR analysis. N = 4 in A, and N = 3 in D and E. Representative blots are shown in B, C, and D from at least three separate experiments. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. AECII = type II alveolar epithelial cell; ND = not determined.