Figure 5

The effect of RNH1 on ferroptosis-associated genes in PBMCs challenged with LPS. Isolated PBMCs were exposed to 50 ng/ml LPS in the presence or absence of 640 U/ml RNH1. Unstimulated cells were used as neg. ctrl. Presented are (A) ACSL1, (B) HMOX1, (C) FTH1, and (D) HAMP relative mRNA expression levels measured using quantitative real-time PCR after stimulation for 4 h (n = 8); (E) HAMP protein concentration quantified using ELISA after 4, 8, 16, and 24 h (n = 3); (F) the correlation between TNFα and HAMP protein concentrations at 4 h (n = 3); and (G) RNase 5 protein concentration quantified using ELISA after 4 h (n = 10). Data are represented as column bar graphs of the mean ± SEM or XY dot plots with the regression line. Repeated-measures one-way ANOVA followed by Tukey’s test or simple linear regression were used for statistical analysis. *p < 0.05; **p < 0.01; ***p < 0.001; RNH1, ribonuclease inhibitor 1; LPS, lipopolysaccharide; PBMCs, peripheral blood mononuclear cells; ACSL1, long-chain acyl-CoA synthetase 1; HMOX1, heme oxygenase 1; FTH1, ferritin heavy chain 1; HAMP, hepcidin, RNase 5, ribonuclease 5.