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. 2020 Mar 22;9(3):775. doi: 10.3390/cells9030775

Figure 3.

Figure 3

Pirfenidone inhibits Extracellular Matrix Protein (ECM) production. (A–G) Intestinal fibroblasts were treated with increasing (maximum 2 mg/mL) concentrations of pirfenidone for 72 h. Pirfenidone suppressed COL1A1, COL3A1, COL4A1, COL6A1, FN1, and ELN mRNA levels, but not ACTA2 levels (* p < 0.05, n = 3). (H) Immunofluorescent images showing that pirfenidone dose-dependently suppressed intracellular levels of collagen I. (I) p-hIFs were exposed for 72 h to pirfenidone (Lanes 1–4) and subsequently cultured in pirfenidone-free medium for 96 h (Lanes 5–8). (J–L) Western blot analysis showed that pirfenidone concentration-dependently and reversibility suppressed collagen I protein levels in p-hIFs. The relative abundance of the tested proteins was normalized to that of Glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ** p < 0.01, *** p < 0.001, **** p < 0.0001; Data are presented as mean ± SEM. Scale bars = 50 μm. Refresh+, refresh medium.