Figure 10.

Effect of CFT073 on hepcidin expression in cultured renal mIMCD-3 cells. mIMCD-3 cells were infected with PFA-fixed CFT073, and the mRNA level of hepcidin was explored at different multiplicities of infection (MOIs; number of bacteria per number of monolayer cells) and different incubation times. (A) The effect of different MOIs on hepcidin mRNA abundance. (B) The time-course effect using an MOI of 5:1. (C) SMAD signaling in mIMCD-3 infected during 24 hours with PFA-fixed CFT073 (MOI of 5:1), was analyzed by western blot. The left panel is a representative western blot of three independent cell cultures, and the right panel is the quantification of pSmad1/5/8 levels. The data were normalized to β-actin. (D) mIMCD-3 cells were incubated with different concentrations of LPS, and the mRNA level of hepcidin was explored by quantitative RT-PCR. All mRNA quantifications were determined by quantitative RT-PCR. Data were normalized by glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-mRNA levels. The results are the mean±SEM of at least six individual mice in each group. *P<0.05; **P<0.03; ***P<0.001. NS, nonsignificant.