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. 2016 May 27;7(28):43731–43745. doi: 10.18632/oncotarget.9667

Figure 3. Effect of NAC and catalase pretreatment on basal and TGF-β1-induced α-SMA, MAPKs and Notch3 expression in IMR-90 fibroblasts.

Figure 3

A. Induction of reactive oxygen species (ROS) by TGF-β1. IMR-90 fibroblasts were treated with TGF-β1 (200 pM, 1 h), with or without NAC or catalase pre-treatment, followed by incubation with 2′,7′-dichlorodihydrofluorescein diacetate (DCF, 10 μM, 15 min). Representative images of DCF fluorescence were shown, and ROS production was quantified and presented relative to that without any treatment as means ± SEM (n = 3; *, P < 0.05). Bar size, 100 μm. B-E. Western analyses of the effect of antioxidant pre-treatment on basal and TGF-β1-induced expression of α-SMA (B), Notch3 (C), phospho-p38 (D), and phospho-JNK1/2 (E). Cells were pretreated with NAC (4 mM, 1 h) or catalase (1000 U/L, 4 h) prior to TGF-β1 (200 pM) stimulation for 48 h (for α-SMA and Notch3) or 1 h (for p38 and JNK1/2). The expression levels were quantified and normalized with their respective controls, and presented relative to that treated with TGF-β1 alone as means ± SEM (n = 3-4; *, P < 0.05). F-G. Expression of phosphorylated ERK1/2, p38, and JNK1/2 in IMR-90 fibroblasts treated with NAC (F, 4 mM) or catalase (G, 1000 U/L) for 0-240 min.