Fig. 8

Effect of GPX4 knock-down on flagellin stimulated NF-κB- reporter activity and endogenous interleukin 8 expression. a Semi-quantitative RTPCR amplification of GPX4 mRNA in cells treated with either a specific GPX4 siRNA (120 pmol) or the negative control. Total RNA was extracted, RTPCR carried out and the products separated by gel electrophoresis. b The intensity of bands corresponding to the amplified GPX4 product was measured using UV Band software, normalised to the housekeeping gene GAPDH and expression then calculated relative to the cells treated with the negative control siRNA. c Quantification of IL8 expression by semi-quantitative RTPCR. Bands corresponding to the amplified IL8 products were normalised to the housekeeping gene GAPDH and expression then calculated relative to the cells treated with the negative control siRNA. d Luciferase activity measured in NF-κB-luc transfectant or TATA-luc transfectant Caco-2 cells after GPX4 siRNA knockdown and stimulation with 100 ng/ml flagellin for 2 h. Luciferase activity was measured in cell extracts, calculated per mg cell protein and expressed relative to the activity in the NF-κB-luciferase cells treated with negative control siRNA. *P < 0.05, **P < 0.01