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. 2018 Jun 27;6(12):e13753. doi: 10.14814/phy2.13753

Figure 8.

Figure 8

Compound 1 treatment of human lung fibroblasts cell cultures increases intracellular cAMP and decreases TGF‐β‐mediated induction of fibrotic gene expression. WI‐38 human lung fibroblasts were seeded on 24‐well plates and treated with increasing concentrations of Compound 1 (1 × 10−10 mol/L to 1 × 10−5 mol/L). After 30 min, cells were stimulated by 1 μmol/L forskolin for 30 min and cell lysates were analyzed for cAMP concentration (A). To analyze fibroblast gene expression, WI‐38 human lung fibroblasts were cultured in media containing 0.5% FBS for 24 h and treated with increasing concentrations of Compound 1 (1 × 10−10 mol/L to 1 × 10−5 mol/L) for 1 h followed by TGF‐β (3 ng/mL) and forskolin (1 μmol/L) for 24 h. Total RNA was extracted from cell lysates, cDNA was amplified, and target gene mRNA was measured. The target gene expression levels of Col1a1 (Type‐1 collagen), Fn (Fibronectin), CTGF (connective tissue growth factor), and PAI‐1 (plasminogen activator inhibitor‐1) was normalized to Glyceraldehyde 3‐phosphate dehydrogenase (GAPDH). Data are reported as mean ± SD with individual data points shown (n = 3 per group). # P ≤ 0.05 versus forskolin group by two‐tailed Williams’ test.