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. 2020 May 15;201(10):1249–1262. doi: 10.1164/rccm.201908-1513OC

Figure 5.

Figure 5.

(A) Hematoxylin and eosin staining of neonatal mouse lungs after 14 days of exposure to normoxia or hyperoxia shows increased alveolar airspaces. Scale bars, 100 μm. (B) Multiplex RNA in situ hybridization (RNA ISH) showing increased expression of Wnt5a (red) coexpressed in cells also expressing fibroblast marker S100a4 (white), with no expression of Wnt5a in cells expressing Sftpc; magenta arrow indicates cells coexpressing S100a4 and Wnt5a. Scale bars, 10 μm. (C) Multiplex RNA ISH demonstrating expression of Wnt target gene Axin2 in both Wnt5A-expressing cells (magenta arrow) and in adjacent Sftpc-expressing alveolar type 2 cells (yellow arrows). Scale bars, 10 μm. (D) Quantification of RNA ISH normalized to cell number shows significantly increased expression of both Wnt5a and Axin2 in response to hyperoxia. ***P < 0.001 and ****P < 0.0001. White boxes in B and C denote area of inset. Data are representative of ≥3 biological replicates for each condition, with 3 technical replicates for histology.