Fig. 2. Effects of uranyl acetate on endogenous H2S generation, H2S synthesizing activity, and mRNA and protein expression of CBS and CSE in normal rat kidney proximal cells (NRK-52E). After the kidney cells were incubated with different concentrations of uranyl acetate (200, 400 or 800 μM) for 24 h, the H2S content in the cell culture supernatant (A) and the H2S synthesizing activity in the kidney cells (B) were tested as described in the “Materials and methods” section. Representative western blots (C) and group data (D) showed CBS and CSE protein abundance in the kidney cells. β-actin was used as the loading control. The CBS and CSE mRNA expression in the kidney cells was determined by real-time PCR (E), and crossing threshold values were normalized to β-actin mRNA expression. Values are expressed as the mean ± SE (n = 5). An asterisk represents a significant difference between uranium-treated and untreated cells (one-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001).
