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. 2019 Nov 21;10:1386. doi: 10.3389/fphar.2019.01386

Figure 2.

Figure 2

Effects of choline and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) on transforming growth factor beta 1 (TGF-β1)-induced collagen secretion. (A) Effects of different concentrations of choline (1, 5, 10 mM) on protein level of collagen I. ***p < 0.001 vs. Ctrl, ## p < 0.01 vs. TGF-β1, n = 5. (B) Immunofluorescence for collagen I in cardiac fibroblasts (CF). Images were obtained using fluorescence microscopy. Blue fluorescence indicates 4′,6-diamidino-2-phenylindole, green fluorescence indicates collagen I, scale bars: 50 μm. *p < 0.05 vs. Ctrl, # p < 0.05 vs. TGF-β1, n = 3. (C) Effect of choline on collagen I protein level in the different experimental groups. *p < 0.05 vs. Ctrl, ## p < 0.01 vs. TGF-β1, && p < 0.01 vs. choline, n = 7. (D) Effect of choline on collagen III protein level in the different experimental groups. *p < 0.05 vs. Ctrl, # p < 0.05 vs. TGF-β1, & p < 0.05 vs. choline, n = 6. (E) Effects of choline (1 mM) and 4-DAMP (3 nM) on protein level of M3 muscarinic acetylcholine receptor (M3R) in TGF-β1-treated CF. *p < 0.05 vs. Ctrl, # p < 0.05 vs. TGF-β1, n = 5. (F) Protein level of M3R after transfecting with three fragments of M3R-siRNA. *p < 0.05 vs. Ctrl, n = 4. (G) Protein level of collagen I after transfecting M3R-siRNA-3. *p < 0.05 vs. Ctrl, n = 5.