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. 2020 Oct 15;5(20):e134464. doi: 10.1172/jci.insight.134464

Figure 2. TRPV4 and aquaporin-5 expression in mouse lungs.

Figure 2

(A) GFP staining (green) by fluorescence-coupled GFP-specific antibodies in lung cryosections of TRPV4EGFP reporter mice reveals expression of TRPV4 in cells of the lung endothelium (EN) as well as in the bronchial (BE) and alveolar epithelium (AE). Nuclei staining was performed with Hoechst dye (blue). Scale bar: 10 μm (right); 20 μm (middle); 50 μm (left). (B) Lung cryosections from TRPV4EGFP– reporter mice were stained with fluorescence-coupled antisera directed against GFP and aquaporin-5 (AQP-5). Confocal images were obtained after excitation at 488 nm (for EGFP, left top, green) or after excitation at 561 nm (for AQP-5, left bottom, red). Both images were merged (right). Nuclei staining was performed with Hoechst dye (blue). A, alveolus; B, bronchus; V, vasculature. The inset shows the bottom boxed region in at higher magnification. Scale bar: 10 μm (inset); 20 μm. (C) TRPV4 mRNA quantification in lung cells using NanoString technology. ATII, alveolar type II cells; EC, endothelial cells; PASMC, precapillary arterial smooth muscle cells; pmLF, primary murine lung fibroblasts. Data represent mean ± SEM from at least 3 independent cell isolations.