Fig. 6
OMV-mediated RNase1 mRNA repression depends on p38 signaling. HULEC-5a were pretreated for 1 h with 10 µM p38i (SB202190) followed by 16 h stimulation with SEC-purified OMVs from Ec, Kp and Sal (MOV1000) as well as TNF-α (10 ng/ml), treated with DMSO as solvent control (Ctrl-DMSO) or left untreated as control (Ctrl). mRNA expression of A) RNase1 and C) CXCL10 was determined by qPCR, normalized to RPS18 and the respective Ctrl. n = 3–5, line at mean, Two-way ANOVA with Tukey‘s multiple comparison test. B) RNase1 protein release from 16 h stimulated HULEC-5a was measured by ELISA, depicted as x-fold protein relative to Ctrl. D) HULEC-5a were pretreated with p38i (10 µM for 1 h) followed by 180 min KpOMV stimulation (MOV1000). Expression and phosphorylation of p38 and STAT1 were determined by Western Blot. β-actin served as a loading control. One representative result is shown. A, C) n = 4, line at mean; Two-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. B) n = 3, line at mean; One-way ANOVA with Tukey ‘s multiple comparison test. *p < 0.05