FIGURE 5.

CnAβ regulates ROS generation. A, relative expression of CnAα and CnAβ mRNA was determined in WT kidney fibroblasts transfected with scrambled or CnAβ-targeting (si-CnAβ) oligonucleotides by qRT-PCR. Data shown are the mean ± S.E. (error bars) of triplicate samples. **, p < 0.01 compared with scrambled. B, Cn enzyme activity in si-CnAβ- or scrambled-transfected WT kidney fibroblasts exposed to NG or HG for 48 h was determined by an in vitro Cn assay. Data shown are the mean ± S.E. of 8 replicates/group. *, p < 0.05 compared with scrambled NG; #, p < 0.05 compared with scrambled HG. C, hypertrophy was measured in scrambled and si-CnAβ-transfected WT kidney fibroblasts by assessing the protein/DNA ratio. Data shown are the mean ± S.E. of 8 replicates/group normalized to scrambled NG. **, p < 0.01 compared with NG. D, ROS generation in scrambled or si-CnAβ-transfected WT kidney fibroblasts exposed to NG or HG for 48 h was examined by Amplex Red. Data shown are the mean ± S.E. of 8 replicates/group normalized to scrambled NG. *, p < 0.05 compared with NG. #, p < 0.05 compared with NG WT. Nox2 (E) and Nox4 (F) mRNA expression in scrambled or si-CnAβ-transfected WT fibroblasts were examined by qRT-PCR. Data shown are the mean ± S.E. of triplicate samples/group normalized to scrambled NG. **, p < 0.01; #, p < 0.05 compared with scrambled NG. ALU, arbitrary light units.