Cathepsin B Regulates Collagen Expression by Fibroblasts via Prolonging TLR2/NF-κB Activation

Figure 3

The effects of CatB on NF-κB activation and oxidative damage in BJ fibroblasts after challenge with P.g. LPS. ((a), (b)) The effect of Bay 11-7082 (Bay, 20 μM, 1 h before P.g. LPS challenge) and SN50 (20 μM, 1 h before P.g. LPS challenge) on the expression of collagen III (a) and collagen IV (b) at 48 h after challenge with P.g. LPS (1 μg/mL). Each column and bar represent the mean ± SEM ( each). The asterisks indicate a statistically significant difference from the value in unchallenged cells (, ). The crosses indicate a statistically significant difference from the value in P.g. LPS-challenged cells without pretreatment with Bay 11-7082 (, ). (c) The time course of the protein expression of CatB in BJ fibroblasts after challenge with P.g. LPS (1 μg/mL). (d) The quantitative analyses of the immunoblotting for CatB. Each column and bar represent the mean ± SEM (, each). The asterisks indicate a statistically significant difference from the value in the unchallenged cells (). (e) The time course of the protein expression of IκBα in BJ fibroblasts after challenge with P.g. LPS (1 μg/mL). (f) The quantitative analyses of the immunoblotting for IκBα. Each column and bar represent the mean ± SEM (, each). The asterisks indicate a statistically significant difference from the value in the unchallenged cells (). The crosses indicate a statistically significant difference from the value in 48 h P.g. LPS-challenged cells without pretreatment with CA-074Me (). (g) The immunofluorescent CLSM images of p65 (green) in BJ fibroblasts after 48 h challenged with P.g. LPS or pretreatment with CA-074Me. Scale bar = 10 μm. (h) The amount of 8-OHdG in BJ fibroblasts 48 h after challenge with P.g. LPS. Each column and bar represent the mean ± SEM (, each). The asterisks indicate a statistically significant difference from the value in the unchallenged cells (). The crosses indicate a statistically significant difference from the value in 48 h P.g. LPS-challenged cells without pretreatment with CA-074Me (). (i) The immunofluorescent CLSM images of 8-OHdG (green) in BJ fibroblasts after 48 h challenged with P.g. LPS or pretreatment with CA-074Me. Scale bar = 10 μm. (j) The protein expression of TLR2 in BJ fibroblasts 48 h after challenge with P.g. LPS (1 μg/mL). (k) The quantitative analyses of the immunoblotting for TLR2 in (j). Each column and bar represent the mean ± SEM (, each). The asterisks indicate a statistically significant difference from the value in the unchallenged cells (). The crosses indicate a statistically significant difference from the value in 48 h P.g. LPS-challenged cells without pretreatment with CA-074Me ().

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)