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. 2022 Apr 27;11(9):1477. doi: 10.3390/cells11091477

Figure 8.

Figure 8

Effect of MDL-800 on TGF-β1-induced extracellular matrix protein expression in HK2 cells. (A) HK2 cells were treated with vehicle (Veh) or TGF-β1 (10 ng/mL), with or without MDL-800 at indicated doses (10, 25, or 50 μM). After 24h of treatment, cell proliferation was measured by XTT assay. Data are expressed as mean ± SD for three independent experiments in triplicates. (B) Representative Western blot for α-SMA, type I collagen, fibronectin, and connective tissue growth factor (CTGF) from HK-2 cells treated with Veh or TGF-β1 (10 ng/mL), with or without MDL-800 at indicated doses (10, 25, or 50 μM). Treatment with TGF-β1 (10 ng/mL) over 24 h increased the expression of extracellular matrix protein markers. The expression of α-SMA, type I collagen, fibronectin, and CTGF decreased after MDL-800 treatment in a dose-dependent manner. The bar graph shows the densitometric quantification presented as the relative ratio of each protein to GAPDH. Data are presented as mean ± SD. (C) Representative Western blot for p-Smad2 and p-Smad3 from HK-2 cells treated with Veh or TGF-β1 (10 ng/mL), with or without MDL-800 at indicated doses (10, 25, or 50 μM). Treatment with TGF-β1 (10 ng/mL) over 24 h increased the expression of p-Smad2 and p-Smad3. The expression of p-Smad2 and p-Smad3 decreased in a dose-dependent manner after MDL-800 treatment. The bar graph shows the densitometric quantification presented as the relative ratio of each protein to Smad2/3. Data are presented as mean ± SD. ***, p < 0.001 versus Veh or MDL; ###, p < 0.001 versus TGF-β1 (10 ng/mL). Veh, vehicle; MDL, MDL-800; TGF-β1, transforming growth factor-β1; α-SMA, α-smooth muscle actin; CTGF, connective tissue growth factor.