Fig. 4

The effects of LE in TGF-β1-stimulated NPDFs for fibroblast differentiation and ECM production. A NPDFs with or without TGF-β1 stimulation (20 ng/mL) were treated with LE (200, 500, and 1000 μg/mL) at an increasing concentration, and the mRNA expression levels of α-SMA (i), FBN (ii), and type I collagen (iii) were tested by RT-PCR. B LE (25, 100, 300, and 1000 μg/mL) was used to treat NPDFs with or without the activation of TGF-β1 (20 ng/mL). Western blotting was carried out to analyze the protein expressions of α-SMA and FBN, and the results were analyzed by ImageJ software after normalization based on GAPDH. The relative differences between the two groups were statistically analyzed. C IF combined with confocal laser microscopy was used to examine the immunofluorescent expression of α-SMA (red fluorescent area indicated by the white arrow) and FBN (green fluorescent area indicated by the white arrow) in NPDFs treated with TGF-β1 (20 ng/mL) alone, LE (500 μg/mL) alone, and the combined TGF-β1 (20 ng/mL) and LE (500 μg/mL)