Figure 6
The effect of RNH1 on the regulation of LPS-induced relative mRNA expression of genes associated with non-canonical inflammasome activation and downstream signaling in PBMCs. 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) GBP1, (B) GBP5, and (C) CASP5 measured using quantitative real-time PCR (n = 8); (D) pro-CASP5 protein expression analyzed using Western blot after 4, 8, 16, and 24 h (n = 3); (E) relative GSDMD mRNA expression after 4 h (n = 8); and (F) IL-1β protein concentrations in PBMC supernatants quantified using ELISA after stimulation for 4, 8, 16, and 24 h (n = 3). Data are represented as column bar graphs of the mean ± SEM. Two-way ANOVA or repeated-measures ANOVA followed by Tukey’s test were used for multiple comparisons. *p < 0.05; **p < 0.01; RNH1, ribonuclease inhibitor 1; LPS, lipopolysaccharide; PBMCs, peripheral blood mononuclear cells; GBP1, guanylate-binding protein 1; GBP5, guanylate-binding protein 5; CASP5, caspase-5; GSDMD, gasdermin D; IL-1β, interleukin-1 β.
