Fig. 3

OMVs repress endothelial RNase1 and induce proinflammatory EC activation in a TLR4-dependent manner. SEC-purified OMVs from Ec, Kp and Sal were pretreated for 1 h with 20 µg/ml Polymyxin B (PB) and further used for stimulation of HULEC-5a (MOV₁₀₀₀) for 16 h, as well as TNF-α (10 ng/ml), LPS from Ec (LPS_Ec) or Sal (LPS_Sal) (100 ng/ml) or left untreated as control (Ctrl). mRNA expression of A) RNase1, C) CXCL8, E) ICAM-1 and F) CXCL10 was determined by qPCR, normalized to RPS18 and Ctrl. n = 3–5, line at mean, Two-way ANOVA with Tukey ‘s multiple comparison test. B) RNase1 protein release and D) CXCL8 protein release from supernatants of 16 h stimulated HULEC-5a was measured by ELISA, depicted as x-fold protein relative to Ctrl. n = 4, line at mean, One-way ANOVA with Tukey ‘s multiple comparison test. * compared to corresponding Ctrl, # as indicated. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. #p < 0.05, ##p < 0.01, ####p < 0.0001